最新elisa實(shí)驗(yàn)的準(zhǔn)備工作發(fā)布

<p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">在常見的實(shí)驗(yàn)中，</font><strong>elisa<font face="宋體">試劑盒</font></strong><font face="宋體">測(cè)定過程為試劑準(zhǔn)備工作，加樣，洗滌，保溫等正常的實(shí)驗(yàn)流程，一次好的準(zhǔn)備工作可以使測(cè)定中試劑發(fā)揮重要的作用，也可以因?yàn)橐粋€(gè)小步驟而發(fā)生多種測(cè)定結(jié)果。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">以下是我司為大家提供整理的</font>elisa<font face="宋體">試劑新實(shí)驗(yàn)準(zhǔn)備要點(diǎn)，僅供大家參考，希望幫大家解決檢測(cè)上的疑問！</font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">試劑的準(zhǔn)備</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">按試劑盒說明書的要求準(zhǔn)備實(shí)驗(yàn)中需用的試劑。</font>ELISA<font face="宋體">中用的蒸餾水或去離子水，包括用于洗滌的，應(yīng)為新鮮的和高質(zhì)量的。自配的緩沖液應(yīng)用</font><font face="Calibri">pH</font><font face="宋體">計(jì)測(cè)量較正。從冰箱中取出的試驗(yàn)用試劑應(yīng)待溫度與室溫平衡后使用。試劑盒中本次試驗(yàn)不需用的部分應(yīng)及時(shí)放回冰箱保存。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">加</span><span style="font-family: 宋體; font-size: 9pt;">&nbsp;&nbsp;&nbsp;&nbsp;</span><span style="font-family: 宋體; font-size: 9pt;">樣</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">在</font>ELISA<font face="宋體">中一般有</font><font face="Calibri">3</font><font face="宋體">次加樣步聚，即加標(biāo)本，加酶結(jié)合物，加底物。加樣時(shí)應(yīng)將所加物加在</font><font face="Calibri">LEISA</font><font face="宋體">板孔的底部，避免加在孔壁上部，并注意不可濺出，不可產(chǎn)生氣泡。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">加標(biāo)本一般用微量加樣器，按規(guī)定的量加入板孔中。每次加標(biāo)本應(yīng)更換吸嘴，以免發(fā)生交叉污染，也可用一次性的定量塑料管加樣。</span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">有此測(cè)定（如間接法</font>ELISA<font face="宋體">）需用稀釋的血清，可在試管中按規(guī)定的稀釋度稀釋后再加樣。也可在板孔中加入稀釋液，再在其中加入血清標(biāo)本，然后在微型震蕩器上震蕩</font><font face="Calibri">1</font><font face="宋體">分鐘以保證混和。加酶結(jié)合物應(yīng)用液和底物應(yīng)用液時(shí)可用定量多道加液器，使加液過程迅速完成。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">保</span><span style="font-family: 宋體; font-size: 9pt;">&nbsp;&nbsp;&nbsp;&nbsp;</span><span style="font-family: 宋體; font-size: 9pt;">溫</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">在</font>ELISA<font face="宋體">中一般有兩次抗原抗體反應(yīng)，即加標(biāo)本和加酶結(jié)合物后?？乖贵w反應(yīng)的完成需要有一定的溫度和時(shí)間，這一保溫過程稱為溫育（</font><font face="Calibri">incubation</font><font face="宋體">），有人稱之為孵育，在</font><font face="Calibri">ELISA</font><font face="宋體">中似不恰當(dāng)。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">elisa<font face="宋體">試劑盒屬固相免疫測(cè)定，抗原、抗體的結(jié)合只在固相表面上發(fā)生。以抗體包被的夾心法為例，加入板孔中的標(biāo)本，其中的抗原并不是都有均等的和固相抗結(jié)合的機(jī)會(huì)，只有貼近孔壁的一層溶液中的抗原直接與抗體接觸。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">這是一個(gè)逐步平衡的過程，因此需經(jīng)擴(kuò)散才能達(dá)到反應(yīng)的終點(diǎn)。在其后加入的酶標(biāo)記抗體與固相抗原的結(jié)合也同樣如此。</span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">洗</span><span style="font-family: 宋體; font-size: 9pt;">&nbsp;&nbsp;&nbsp;&nbsp;</span><span style="font-family: 宋體; font-size: 9pt;">滌：</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">洗滌在</font>ELISA<font face="宋體">過程中雖不是一個(gè)反應(yīng)步驟，但卻也決定著實(shí)驗(yàn)的成敗。</font><font face="Calibri">ELSIA</font><font face="宋體">就是靠洗滌來達(dá)到分離游離的和結(jié)合的酶標(biāo)記物的目的。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">通過洗滌以清除殘留在板孔中沒能與固相抗原或抗體結(jié)合的物質(zhì)，以及在反應(yīng)過程中非特異性地吸附于固相載體的干擾物質(zhì)。聚苯乙烯等塑料對(duì)蛋白質(zhì)的吸附是普遍性的，而在洗滌時(shí)又應(yīng)把這種非特異性吸附的干擾物質(zhì)洗滌下來。</span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">可以說在</font>ELISA<font face="宋體">操作中，洗滌是主要的關(guān)鍵技術(shù)，應(yīng)引起操作者的高度重視，操作者應(yīng)嚴(yán)格按要求洗滌，不得馬虎。洗滌的方式除某些</font><font face="Calibri">elisa</font><font face="宋體">試劑盒儀器配有特殊的自動(dòng)洗滌儀外，手工操作有浸泡式和流水沖洗式兩種</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">顯色和比色</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">（</font>1<font face="宋體">） </font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">顯色</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">顯色是</font>ELISA<font face="宋體">中的后一步溫育反應(yīng)，此時(shí)酶催化無色的底物生成有色的產(chǎn)物。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">（</font>2<font face="宋體">）</font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體"> 比色</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">比色前應(yīng)先用潔凈的吸水紙拭干板底附著的液體，然后將板正確放入酶標(biāo)比色儀的比色架中。</span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>