ELISAS實驗血清的制備

<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;">①&nbsp;&nbsp;<strong><font face="Calibri">ELISA</font><font face="宋體">試劑盒</font></strong><font face="宋體">所需的抗原量 一般</font><font face="Calibri">1-10mg/mL</font><font face="宋體">。</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;">② 免疫途徑 抗原的注射方法有皮下注射、肌肉注射、腹膜內(nèi)注射、靜脈注射和淋巴結(jié)注射等。其中以靜脈注射最為簡單方便，不需要佐劑乳化，產(chǎn)生抗體的高峰值出現(xiàn)較早，一般在一周左右，注射后一周即可采血測試效價。免疫過程中應(yīng)注意：<font face="Calibri">a.</font><font face="宋體">佐劑只能現(xiàn)配現(xiàn)用：</font><font face="Calibri">b.</font><font face="宋體">采血時間不宜拖得太長。因為免疫時間越長，產(chǎn)生交叉反應(yīng)的抗體也越多。</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;">③&nbsp;<strong><font face="Calibri">ELISA</font><font face="宋體">試劑盒</font></strong><font face="宋體">抗血清的分離 當(dāng)?shù)玫綕M意的效價時，即可進(jìn)行取血。將收集到的血液成斜面放置在</font><font face="Calibri">37</font><font face="宋體">℃、溫箱中</font><font face="Calibri">2h</font><font face="宋體">。然后置于</font><font face="Calibri">4</font><font face="宋體">℃冰箱過一夜。次日用滴管吸取抗血清，用離心法（</font><font face="Calibri">2000-3000r/min,20min</font><font face="宋體">）去掉沉淀。然后測定效價、鑒定純度。純度用紫外分光光度計或</font><font face="Calibri">Folin</font><font face="宋體">氏比色法進(jìn)行測定。抗血清保存于</font><font face="Calibri">-25</font><font face="宋體">℃低溫處。</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;">④抗血清的效價測定 測定的方法一般有兩種，即試管沉淀法和免疫雙擴(kuò)散法。對于免疫酶技術(shù)使用的抗血清，必須具有一定的效價，試管沉淀法應(yīng)不低于<font face="Calibri">1:2</font><font face="宋體">，免疫雙擴(kuò)散法不低于</font><font face="Calibri">1:2</font><font face="宋體">，且要求單價抗血清。</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">試劑盒捕食者的收集和準(zhǔn)備 主要方法有</font><font face="Calibri">3</font><font face="宋體">個：①將胃內(nèi)含物擠在質(zhì)量好的濾紙上，但蛋白質(zhì)易與纖維素結(jié)合，若保存過久則難以洗脫：②將待檢查的捕食封閉在明膠膠囊內(nèi)，并加干燥劑，以使蟲體與膠囊分離：③盡可能快地將采集到的捕食者儲存于低溫下（如</font><font face="Calibri">-40</font><font face="宋體">℃）。前兩種方法適于郵寄和長途運輸，后一種方法能保存較長的時間。所有的方法都需保持干燥。對較大的捕食者，可將消化道分離，像蟻、蝸牛以及蜘蛛等小型動物則整個蟲體。（</font><font face="Calibri">4</font><font face="宋體">）酶聯(lián)抗體的制備 按戊二醛簡易法進(jìn)行。</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>