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class="content"><p><span>需求數(shù)量：</span>0</p><p><span>價格要求：</span>面議</p><p><span>所在地：</span>上海市</p><p><span>包裝要求：</span><span style="color: #f30;"></span></p><p><span>產(chǎn)品關(guān)鍵詞：</span>上海四色免疫組化染色服務(wù)</p><p><span>***更新：</span>2020-06-09 08:19:56</p><p><span id="click" >瀏覽次數(shù)：0次</span></p><a rel="nofollow" class="button">聯(lián)系我們</a></div></div><div id="exphov8" class="rightbox"><h3>公司基本資料信息</h3><p style="border:0;"><a rel="nofollow">上海朝瑞生物科技有限公司</a></p><p><span>聯(lián)系人：</span>仲先生</p><p><span>郵箱: </span>scienceladder@163.com</p><p><span>電話: </span>18621867065</p><p><span>傳真: </span>021_</p><p><span>網(wǎng)址: </span></p><p><span>手機: </span>021-54133071</p><p><span>地址: </span>上海市松江區(qū)廣富林路697號昂立大廈2113室</p><p>[當前離線] <a rel="nofollow"> [加為商友] </a><a rel="nofollow"> [發(fā)送信件] </a></p></div></div><div id="vta3qnr" class="page-proshow02"><p class="title">詳細說明</p><div id="unfceln" class="box"><p><p> 6）滴加通用型IgG抗體-Fab段-HRP多聚體，室溫/37℃孵育30分鐘-1h，PBS/TBS沖洗，3分鐘×5次。7）應(yīng)用DAB溶液顯色。8）蒸餾水沖洗、復(fù)染、脫水、透明、封片。三、免疫熒光技術(shù)（略）1、酶免疫組化的關(guān)鍵環(huán)節(jié)1）標本固定固定的目的是①防止標本從玻片上脫落；②除去妨礙抗原-抗體結(jié)合的類脂，使抗原抗體結(jié)合物易于獲得良好的染色結(jié)果；③固定的標本易于保存。固定劑的選擇一般用4%多聚甲醛，但睪丸組織、眼可能要選用Bouin’s液或mDF液效果較好。2）脫水、石蠟包埋和制片脫水用梯度乙醇（由低到高）充分脫水、對組織要完全浸蠟，上海四色免疫組化染色服務(wù)、切片時刀片要干凈和鋒利。否則，容易裂片和脫片等。3）脫蠟和水化這是為了后面的抗體等試劑能夠充分與組織中抗原等結(jié)合反應(yīng)。脫蠟可以先60℃20min，然后立即二甲苯1-3分別10min（這個時間是由二甲苯新鮮程度和室溫等綜合決定的），但當天制好的切片一般先60℃3-4h，上海四色免疫組化染色服務(wù)。水化用梯度乙醇（由高到低）。若脫蠟和水化不全易出現(xiàn)局灶性反應(yīng)和浸洗不全，而產(chǎn)生非特異性背景著色，上海四色免疫組化染色服務(wù)。4）抗原修復(fù)由于組織中部分抗原在甲醛或多聚甲醛固定過程中，發(fā)生了蛋白之間交聯(lián)及醛基的封閉作用，從而失去抗原性；通過抗原修復(fù)，使得細胞內(nèi)抗原決定族重新暴露。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海四色免疫組化染色服務(wù)" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899754550782/2606693.png"></div><p> ·若標本自發(fā)熒光對照和特異性對照呈無熒光或弱熒光，陽性對照和待檢標本呈強熒光，則為特異性陽性染色。?直接免疫熒光法的注意事項(續(xù))·一般標本在高壓汞燈下照射超過3min，就有熒光減弱現(xiàn)象；·經(jīng)熒光染色的標本比較好在當天觀察，隨著時間的延長，熒光強度會逐漸下降。(2)間接法又稱為熒光抗-抗體法?需要兩種抗體參與，即一抗和二抗(熒光素標記)。一抗對標本中的抗原來說起抗體的作用，但對熒光標記的二抗來說又起著抗原作用。–可用來檢測標本中未知抗原，也可檢測血清中未知抗體。?間接免疫熒光法操作步驟·標本的處理及非特異染色的封閉同直接法；·一抗染色：–加未標記的特異性抗體(通常1:100稀釋，用)，37?C作用30min或4?C過夜?！BST漂洗5min×3次(震蕩漂洗)；?間接免疫熒光法操作步驟(續(xù))·加熒光標記的二抗抗體，37?C濕盒避光作用30min?！BST避光漂洗5min×3次(例如包上錫紙，在搖床上漂洗)；·甘油緩沖液封片·鏡檢·優(yōu)點：敏感性較高，比直接法高10倍左右；制備一種熒光標記抗體，可應(yīng)用于多種一抗；·缺點：是參加反應(yīng)的因子較多，產(chǎn)生非特異性染色的機會增多。?間接免疫熒光法的注意事項·熒光染色后一般在1h內(nèi)完成觀察，或于4℃保存4h。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海四色免疫組化染色服務(wù)" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748701172/8129573.png"></div><p> ⑴抗原有無丟失主要看組織切片的新鮮程度，一般切片室溫保存超過3-6個月，可能切片內(nèi)的抗原丟失很嚴重（有文獻支持），此時可以通過重新用石蠟塊切片來進一步驗證（蠟塊需要低溫保存）。⑵石蠟切片在制作過程中可能因醛基對抗原決定族的封閉，這需要通過抗原修復(fù)來充分暴露，從而增加抗原抗體結(jié)合反應(yīng)，提高陽性率，我一般用6min*4次中火微波抗原修復(fù)，用枸櫞酸鈉緩沖液。⑶組織切片中本身抗原含量的多少？這方面我有教訓的，我做胎鼠睪丸間質(zhì)細胞鑒定，用1：200一抗效果很好；但我用1：200一抗孵育成年睪丸間質(zhì)細胞檢測，幾乎呈陰性，后來證實是因為兩者抗原含量相差甚遠，則一抗也要適當提高濃度。2、其次，檢查抗體有無選擇錯誤和抗體孵育條件是否不適。⑴一抗選擇單克隆抗體易出現(xiàn)陰性結(jié)果，因為靈敏度低但特異性好；一抗的speciesreactivity中無檢測組織的種屬，這是比較常見的錯誤；一抗選擇rat，而二抗是抗mouse/rabbit等均有可能出現(xiàn)陰性結(jié)果。⑵抗體孵育時間過短，容易導致陰性結(jié)果。一般一抗我建議4℃孵育過夜和37℃復(fù)溫45min；二抗我一般37℃30min。我不喜歡參照二抗染色試劑盒說明書。⑶抗體濃度過低。這是陰性結(jié)果的**可能原因。必須提高稀釋度。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海四色免疫組化染色服務(wù)" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750107422/8042334.png"></div></p><br /><p>文章來源地址: http://www.cdcfah.com/cp/199962.html</p></div></div></div></div></div><div id="wlnp3ax" class="page-right twoColRt"><div id="jx8czl3" class="box01 margin10 clearfix"><h3><a 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