ELISA检测方法步骤

<p>&nbsp;<font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">　　</font><font face="Calibri" style="font-size: 10.5pt;">ELISA</font><font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">是一种用于检测和定量蛋白质、抗体、抗原的免疫学方法，广泛应用于生物医学研究和诊断中。然而，对于刚接触</font><font face="Calibri" style="font-size: 10.5pt;">ELISA</font><font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">新手来说，复杂的实验步骤和细节常常让他们感到困惑。接下来，我们将逐步讲解</font><font face="Calibri" style="font-size: 10.5pt;">ELISA</font><font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">的操作步骤。</font></p> <p class="MsoNormal" style="text-align: center;"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;<img src="/images/upload/Image/5(47).png" alt="酶联生物ELISA试剂盒" width="500" height="375" /></span></p> <h3><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　实验步骤</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></h3> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　</font><font face="Calibri">&nbsp;</font><strong><font face="Calibri">一、</font><font face="宋体">实验试剂准备</font></strong></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　1.抗原</font><font face="Calibri">/</font><font face="宋体">抗体</font><font face="Calibri">:</font><font face="宋体">根据实验目的和</font><font face="Calibri">ELISA</font><font face="宋体">类型</font><font face="Calibri">(</font><font face="宋体">例如夹心法、间接法、竞争法</font><font face="Calibri">)</font><font face="宋体">，选择合适的抗原或抗体。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　2.</font><font face="Calibri">96</font><font face="宋体">孔板</font><font face="Calibri">:</font><font face="宋体">高结合力的聚苯乙烯微孔板，用于吸附抗原或抗体。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　3.酶标二抗</font><font face="Calibri">:</font><font face="宋体">与目标抗体</font><font face="Calibri">/</font><font face="宋体">抗原结合的酶标二抗，例如辣根过氧化物酶</font><font face="Calibri">(HRP)</font><font face="宋体">或碱性磷酸酶</font><font face="Calibri">(AP)</font><font face="宋体">标记的二抗。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　4.封闭液</font><font face="Calibri">:</font><font face="宋体">用于封闭微孔板未结合位点，减少非特异性结合。常用</font><font face="Calibri">5%BSA</font><font face="宋体">（牛血清白蛋白）</font><font face="Calibri">PBS</font><font face="宋体">溶液或脱脂奶粉</font><font face="Calibri">PBS</font><font face="宋体">溶液。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　5.酶底物</font><font face="Calibri">:</font><font face="宋体">用于与酶标二抗上的酶反应，产生可检测的信号。</font><font face="Calibri">HRP</font><font face="宋体">常用</font><font face="Calibri">TMB</font><font face="宋体">（</font><font face="Calibri">3,3',5,5'-</font><font face="宋体">四甲基联苯胺），</font><font face="Calibri">AP</font><font face="宋体">常用</font><font face="Calibri">pNPP</font><font face="宋体">（对硝基苯磷酸）。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　6.酶反应终止液</font><font face="Calibri">:</font><font face="宋体">用于终止酶促反应。</font><font face="Calibri">HRP</font><font face="宋体">常用</font><font face="Calibri">2N</font><font face="宋体">硫酸溶液。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　7.洗涤液</font><font face="Calibri">:</font><font face="宋体">用于洗涤未结合的抗体或抗原，常用</font><font face="Calibri">0.05%PBST(PBS</font><font face="宋体">溶液</font><font face="Calibri">+0.05%Tween20)</font><font face="宋体">。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　8.样品稀释液</font><font face="Calibri">:</font><font face="宋体">根据样品的不同，选择合适的稀释液。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　9.标准品</font><font face="Calibri">(</font><font face="宋体">可选</font><font face="Calibri">):</font><font face="宋体">如果需要进行定量分析，则需要准备一系列已知浓度的标准品，用于绘制标准曲线。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">二、实验步骤</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"> </span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"> </span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　1.包被抗体</font><font face="Calibri">/</font><font face="宋体">抗原</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　配制合适浓度的抗原或抗体（通常</font><font face="Calibri">1-10&micro;g/mL</font><font face="宋体">），浓度需要通过预实验进行确定。用碳酸盐缓冲液（</font><font face="Calibri">pH9.6</font><font face="宋体">）稀释。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　向</font><font face="Calibri">96</font><font face="宋体">孔板中每孔加入合适的体积（例如</font><font face="Calibri">50-200&micro;</font><font face="宋体">）稀释好的抗原或抗体溶液。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　在</font><font face="Calibri">4</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">或</font><font face="Calibri">37</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">孵育过夜或</font><font face="Calibri">2</font><font face="宋体">小时，让抗原</font><font face="Calibri">/</font><font face="宋体">抗体吸附到板子上。孵育时间和温度需要根据具体情况进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　三、洗板</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　用</font><font face="Calibri">PBST</font><font face="宋体">（含</font><font face="Calibri">0.05%Tween-20</font><font face="宋体">的</font><font face="Calibri">PBS</font><font face="宋体">）洗板，重复以下步骤</font><font face="Calibri">3-5</font><font face="宋体">次，或根据需要增加洗涤次数，以确保充分去除未结合的物质：</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　①、弃去孔内液体。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　②、用足量的洗涤液</font><font face="Calibri">(</font><font face="宋体">例如</font><font face="Calibri">200-300&micro;L/</font><font face="宋体">孔</font><font face="Calibri">)</font><font face="宋体">充满每孔。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　③、静置</font><font face="Calibri">1-2</font><font face="宋体">分钟。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　④、弃去洗涤液，并用吸水纸小心拍干板子，避免交叉污染。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">四、封闭</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　每孔加入</font><font face="Calibri">200&micro;L</font><font face="宋体">封闭液，例如</font><font face="Calibri">1%-5%BSA</font><font face="宋体">或脱脂奶粉的</font><font face="Calibri">PBS/PBST</font><font face="宋体">溶液。在室温下孵育</font><font face="Calibri">1-2</font><font face="宋体">小时，或</font><font face="Calibri">4</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">过夜孵育。孵育结束后，进行洗板操作：</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　①、弃去孔内液体。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　②、用洗涤液</font><font face="Calibri">(</font><font face="宋体">例如</font><font face="Calibri">0.05%PBST)</font><font face="宋体">注满每孔。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　③、静置</font><font face="Calibri">1-2</font><font face="宋体">分钟。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　④、弃去洗涤液，并用吸水纸小心拍干板子，避免交叉污染。重复洗板步骤</font><font face="Calibri">3-5</font><font face="宋体">次。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">五、加样品或标准品</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　使用样品稀释液或标准品稀释液（通常与封闭液成分相同）按照合适的稀释梯度（例如</font><font face="Calibri">2</font><font face="宋体">倍、</font><font face="Calibri">3</font><font face="宋体">倍、</font><font face="Calibri">5</font><font face="宋体">倍等）稀释待测样品或标准品。对于定量</font><font face="Calibri">ELISA</font><font face="宋体">，需要准备一系列已知浓度的标准品，并进行梯度稀释，用于绘制标准曲线。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　每孔加入</font><font face="Calibri">100&micro;L</font><font face="宋体">稀释后的样品或标准品。设置空白对照孔，不加样品或标准品，只加稀释液。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　在室温下孵育</font><font face="Calibri">1-2</font><font face="宋体">小时，或</font><font face="Calibri">37</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">孵育</font><font face="Calibri">30-60</font><font face="宋体">分钟。孵育时间和温度需要根据具体的抗原抗体进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">六、加检测抗体（对于夹心法）</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　孵育完样品后，进行洗板操作：</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　①、弃去孔内液体。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　②、用洗涤液</font><font face="Calibri">(</font><font face="宋体">例如</font><font face="Calibri">0.05%PBST)</font><font face="宋体">充满每孔。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　③、静置</font><font face="Calibri">1-2</font><font face="宋体">分钟。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　④、弃去洗涤液，并用吸水纸小心拍干板子，避免交叉污染。重复洗涤步骤</font><font face="Calibri">3-5</font><font face="宋体">次。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　⑤、加入用抗体稀释液</font><font face="Calibri">(</font><font face="宋体">通常与封闭液成分相同</font><font face="Calibri">)</font><font face="宋体">稀释好的检测抗体</font><font face="Calibri">(</font><font face="宋体">结合靶标抗原的酶标抗体</font><font face="Calibri">)</font><font face="宋体">，每孔</font><font face="Calibri">100&micro;L</font><font face="宋体">。抗体的稀释倍数需要根据具体的抗体和实验条件进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　⑥、在室温或</font><font face="Calibri">37</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">下孵育</font><font face="Calibri">1-2</font><font face="宋体">小时。具体孵育时间和温度需要根据具体的抗体进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　⑦、孵育结束后，再次进行洗板操作</font><font face="Calibri">(</font><font face="宋体">步骤同上</font><font face="Calibri">)</font><font face="宋体">。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">七.加酶标二抗</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　使用抗体稀释液（通常与封闭液成分相同）稀释酶标二抗。加入稀释好的酶标二抗（结合检测抗体的酶标抗体），每孔</font><font face="Calibri">100&micro;L</font><font face="宋体">。二抗的稀释倍数需要根据具体的抗体和实验条件进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　在室温或</font><font face="Calibri">37</font><font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">下孵育</font><font face="Calibri">30</font><font face="宋体">分钟至</font><font face="Calibri">1</font><font face="宋体">小时。具体孵育时间和温度需要根据具体的二抗进行优化。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　孵育结束后，进行洗板操作：</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　①、弃去孔内液体。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　②、用洗涤液</font><font face="Calibri">(</font><font face="宋体">例如</font><font face="Calibri">0.05%PBST)</font><font face="宋体">充满每孔。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　③、静置</font><font face="Calibri">1-2</font><font face="宋体">分钟。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　④、弃去洗涤液，并用吸水纸小心拍干板子，避免交叉污染。重复洗板步骤</font><font face="Calibri">3-5</font><font face="宋体">次。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">八.加底物反应</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　根据试剂盒说明书准备酶底物溶液。加入</font><font face="Calibri">100&micro;L</font><font face="宋体">酶底物溶液（如</font><font face="Calibri">TMB</font><font face="宋体">）到每孔，避光反应一段时间，通常为</font><font face="Calibri">5-30</font><font face="宋体">分钟，或直至出现明显的颜色变化</font><font face="Calibri">(</font><font face="宋体">例如，对于</font><font face="Calibri">TMB</font><font face="宋体">底物，颜色会从无色变为蓝色</font><font face="Calibri">)</font><font face="宋体">。密切观察显色情况，并根据试剂盒说明书和标准品的显色情况来确定反应时间。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">九.终止反应</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　根据试剂盒说明书加入终止液，以终止反应，停止底物显色反应。颜色变化取决于所使用的底物。例如，使用</font><font face="Calibri">TMB</font><font face="宋体">底物时，加入</font><font face="Calibri">2N</font><font face="宋体">硫酸后，颜色会从蓝色变为黄色。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　</span><strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">十.测定吸光值</span></strong><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　使用已校准的酶标仪，根据试剂盒说明书选择合适的检测波长和参考波长，读取各孔的吸光值（</font><font face="Calibri">OD</font><font face="宋体">值）。读取吸光值后，需要对数据进行分析，例如绘制标准曲线、计算样品浓度等。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <h3><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　实验注意事项：</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></h3> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　1.重复性：进行足够的重复实验，例如至少三个技术重复（同一份样品在同一块板上进行三次测定）或三个生物学重复（三个不同的样品分别进行测定）。技术重复可以评估实验的精确度，而生物学重复可以评估实验的重复性。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　2.温度控制：在孵育阶段，温度对结果的影响较大。建议使用培养箱进行孵育，以获得更精确的温度控制。如果在室温下孵育，则尽量保持稳定的室温，避免较大的温度波动。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　3.洗涤：每次洗板时，确保洗涤液完全覆盖每个孔。洗涤时，可以轻轻拍打板子边缘，以去除残留液体，或使用洗板机进行洗涤。注意避免交叉污染。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　4.显色时间控制：显色反应时间对结果有很大影响。密切观察显色情况，并根据试剂盒说明书和标准品的显色情况来确定最佳反应时间。当标准品孔出现明显的梯度颜色变化时，即可终止反应。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　5.试剂质量：使用高质量的试剂对于获得可靠的实验结果至关重要。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">　　6.操作规范：严格按照操作规程进行实验，避免人为误差。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　7.标准曲线：对于定量</font><font face="Calibri">ELISA</font><font face="宋体">，标准曲线的质量直接影响样品浓度的计算结果。确保标准曲线线性良好，并选择合适的拟合方法。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体">　　上述是</font><font face="Calibri">ELISA</font><font face="宋体">检测方法步骤，</font><font face="Calibri">ELISA</font><font face="宋体">实验操作包括包被、封闭、加样、洗板、显色等多个步骤。实验完成后，通过标准曲线拟合和计算，可以准确获得样品中的目标分子浓度。在实验中要注意些事项。关于标准曲线拟合和计算，下篇文章继续为您介绍。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>