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多克隆抗體制備|實(shí)驗(yàn)技術(shù)服務(wù)

時(shí)間:2015/5/15閱讀:277
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關(guān)鍵詞:多克隆抗體制備|實(shí)驗(yàn)技術(shù)服務(wù)
簡(jiǎn)介:世界*品牌多克隆抗體制備|實(shí)驗(yàn)技術(shù)服務(wù)原裝,,質(zhì)量保證,*。
多克隆抗體制備|實(shí)驗(yàn)技術(shù)服務(wù)——pCzn1質(zhì)粒+Arctic Express宿主菌低溫表達(dá)體系,。
我們具備豐富的蛋白表達(dá)設(shè)計(jì)經(jīng)驗(yàn)及實(shí)驗(yàn)操作技巧,,承諾客戶不成功不收費(fèi)。我們所有的實(shí)驗(yàn)數(shù)據(jù)真實(shí)可信,,我們提供原始的表達(dá)菌株和克隆質(zhì)粒,,客戶可以依據(jù)我們的實(shí)驗(yàn)報(bào)告重復(fù)我們的實(shí)驗(yàn)結(jié)果。
產(chǎn)品品牌:   http://www.,。,。com/goodsid/fenleiyi/2868603/1.html    
測(cè)序?qū)嶒?yàn)流程:
多克隆抗體的制備步驟
  1) 家兔仰臥于兔架上固定頭部,用紗布固定四肢,。頭部略放低以暴露頸部,。剃毛并消毒皮膚。
   2) 沿頸部中線用手術(shù)刀切開(kāi)皮膚約10cm,,分離皮下結(jié)締組織,,直至暴露出氣管兩側(cè)的胸鎖乳突?。ㄐ⌒牟僮鳎缗龅叫⊙艹鲅?,可用止血鉗止血),。
   3) 用直頭止血鉗分離胸鎖乳突肌與氣管間的頸三角區(qū)疏松組織,,暴露出頸總動(dòng)脈后用彎頭眼科鑷使之游離,,剝離神經(jīng)和結(jié)締組織。
   4) 于動(dòng)脈下套入兩根黑絲線,,分別置于遠(yuǎn)心及近心端,。結(jié)扎遠(yuǎn)心端的絲線。近心端的動(dòng)脈用血管夾夾住,。
   5) 用小拇指墊在血管下,,用尖頭眼科手術(shù)剪在兩根絲線間的動(dòng)脈璧上剪一小口(勿剪斷),插入塑料放血管,。再將近心端的絲線結(jié)扎固定于放血管上,,以防放血管滑脫。
   6) 松開(kāi)止血鉗,,使血液流入容器中,。一般一只家兔可放血100-120ml。
3. 血清的分離
如果用三角瓶或平皿盛血,,將器皿傾斜放置于37oC烘箱2hr,,轉(zhuǎn)移到4oC沉淀,第二天早上用吸管吸取血清,。如果用離心管收集,,37oC烘箱放置2hr,轉(zhuǎn)移到4oC沉淀,,第二天早上離心,,10000RCF,10分鐘,。在血清中加入NaN3至終濃度0.02%,,分裝后-20oC保存
抗血清收獲后,加 1/萬(wàn)硫柳汞或 1/萬(wàn)的疊氮鈉防腐,,或加入等量的中性甘油,,分裝小瓶,置-20℃以下的低溫保存,,數(shù)月至數(shù)年內(nèi)抗體效價(jià)無(wú)明顯變化,。注意避免反復(fù)凍融。也可將抗血清冷凍干燥后保存,。
多克隆抗體的制備一般包括以下幾個(gè)步驟:
1,、制備抗原,。
2、選擇實(shí)驗(yàn)動(dòng)物,。
3,、動(dòng)物免疫。
4,、試取血進(jìn)行測(cè)試,,看看是否成功免疫。
5,、如果成功免疫,,殺死實(shí)驗(yàn)動(dòng)物,采集全部血清,。
6,、純化出抗體。
7,、鑒定抗體,。包括純度以及特異性。
(一)抗原制備
Ig 是免疫球蛋白,,對(duì)異種動(dòng)物而言,,是良好的抗原物質(zhì)??梢源碳ぎ惙N動(dòng)物產(chǎn)生抗 Ig 血清,,
經(jīng)適當(dāng)提取后,產(chǎn)生抗 Ig 抗體,,又叫第二抗體或抗抗體,。在多數(shù)的間接標(biāo)記反應(yīng)中,均需制備
抗 Ig 抗體,。
(二)材料與試劑
1.提取的動(dòng)物 Ig
2.弗氏*佐劑和弗氏不*佐劑
3.青霉素和鏈霉素
4.實(shí)驗(yàn)動(dòng)物 兔
5.其它材料及試劑

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