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src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748427735/7999973.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div><div id="cbif7ac" class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748427735/7999973.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="x3w4a22" class="content"><p><span>需求數(shù)量：</span>0</p><p><span>價(jià)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>天津多色熒光免疫組化分析服務(wù),免疫組化</p><p><span>***更新：</span>2020-07-02 22:19:51</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="cldkh3v" 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(jī): </span>021-54133071</p><p><span>地址: </span>上海市松江區(qū)廣富林路697號(hào)昂立大廈2113室</p><p>[當(dāng)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="2mjqnhy" class="page-proshow02"><p class="title">詳細(xì)說(shuō)明</p><div id="qu3lxof" class="box"><p><p> 在室溫下孵育5分鐘以封閉非特異性的背景染色。（注：孵育不要超過(guò)10分鐘，否則會(huì)導(dǎo)致特異性染色降低。如果一抗的稀釋液中含有5－10%正常羊血清，這一步可以省略。）6.緩沖液洗5min/2次。7.滴加一抗工作液37℃孵育1－2小時(shí)。（具體孵育時(shí)間和溫度由試驗(yàn)者**終決定）8.緩沖液洗5min/2次。9．滴加PrimaryAntibodyEnhancer（增強(qiáng)子），在室溫下孵育20分鐘。10．緩沖液洗5min/2次。11．滴加HRPPolymer（酶標(biāo)二抗），在室溫下孵育30分鐘。（注：HRPPolymer對(duì)光敏感，應(yīng)避免不必要的光暴露并儲(chǔ)存在不透明的小瓶中。）12．緩沖液洗5min/2次。13．向1mlDABPlusSubstrate（或AECPlusSubstrate)中滴加1-2滴DABPlusChromogen（或AECPlusChromogen），混勻后滴加到切片上，孵育3－15分鐘。（具體時(shí)間由染色深淺決定。）14．自來(lái)水充分沖洗，復(fù)染，脫水，透明，封片。免疫組化操作步驟⑴、石蠟切片脫蠟至水。⑵、3%H2O2室溫孵育5-10分鐘，以消除內(nèi)源性過(guò)氧化物酶的活性。⑶、蒸餾水沖洗，PBS浸泡5分鐘x2（如需抗原修復(fù)，可在此步后進(jìn)行）。⑷、5-10%正常山羊血清（PBS稀釋?zhuān)┓忾]，天津多色熒光免疫組化分析服務(wù)，室溫孵育10分鐘，傾去血清，勿洗。滴加一抗工作液，37℃孵育1-2小時(shí)或4℃過(guò)夜，天津多色熒光免疫組化分析服務(wù)。⑸，天津多色熒光免疫組化分析服務(wù)、PBS沖洗，5分鐘x3次。天津多色熒光免疫組化分析服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="天津多色熒光免疫組化分析服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748427735/7999973.png"></div><p> 5、免疫組化技術(shù)掌握與否的鑒定標(biāo)準(zhǔn)是同一切片或不同切片中不同抗原均從摸索濃度或條件而做出優(yōu)良的染色切片。我發(fā)現(xiàn)許多研究生把網(wǎng)上或者說(shuō)明書(shū)摸索的反應(yīng)條件、濃度、方法步驟，重復(fù)運(yùn)用于同一性質(zhì)的切片和同一種抗體，做出來(lái)后就覺(jué)得自己已經(jīng)掌握了免疫組化方法，更換一種抗體后，居然連二抗的種屬來(lái)源都拿錯(cuò)了。失敗往往促進(jìn)你去思考試驗(yàn)原理和過(guò)程，成功有時(shí)也讓自己很自傲。6、免疫組化的應(yīng)用***，是當(dāng)前實(shí)驗(yàn)研究的**重要方法之一。如今發(fā)SCI論文時(shí)，明顯感覺(jué)*靠量化的數(shù)據(jù)來(lái)發(fā)文章很難，加一些形態(tài)學(xué)數(shù)據(jù)或圖片，老外十分歡迎，可能是怕你學(xué)術(shù)造假吧。當(dāng)然也不能做假陽(yáng)性或假陰性結(jié)果。7、實(shí)驗(yàn)方法需要?jiǎng)邮趾蛣?dòng)腦。經(jīng)過(guò)了反復(fù)的動(dòng)手和動(dòng)腦，把理論原理運(yùn)用于實(shí)踐，在把實(shí)踐中發(fā)現(xiàn)的問(wèn)題帶到理論知識(shí)中去解決，**終把理論與實(shí)踐融會(huì)貫通。必要時(shí)去各大論壇或者一些**公司尋求幫助，個(gè)人感覺(jué)上海舜田生物技術(shù)支持不錯(cuò)，服務(wù)質(zhì)量也很好。下面先介紹下免疫組化的概念和常用方法，以便讓大家對(duì)免疫組化更深入地理解和掌握。概念和常用方法介紹1、定義用標(biāo)記的特異性抗體對(duì)組織切片或細(xì)胞標(biāo)本中某些化學(xué)成分的分布和含量進(jìn)行組織和細(xì)胞原位定性、定位或定量研究。天津多色熒光免疫組化分析服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="天津多色熒光免疫組化分析服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899747812500/5201169.png"></div><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(biāo)本固定固定的目的是①防止標(biāo)本從玻片上脫落；②除去妨礙抗原-抗體結(jié)合的類(lèi)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(biāo)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(duì)組織要完全浸蠟、切片時(shí)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(gè)時(shí)間是由二甲苯新鮮程度和室溫等綜合決定的），但當(dāng)天制好的切片一般先60℃3-4h。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。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(guò)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(guò)抗原修復(fù)，使得細(xì)胞內(nèi)抗原決定族重新暴露。</p><p> 在肌肉、皮下、靜脈或腹腔內(nèi)注射；·抗體效價(jià)的檢測(cè)：加強(qiáng)免疫一周后，耳緣靜脈**–抗體效價(jià)測(cè)定可用環(huán)狀沉淀試驗(yàn)、瓊脂雙向擴(kuò)散法等方法。前者抗體效價(jià)在1:4000以上，后者在1:16以上，即可**，取血清進(jìn)一步提純。舉例2：微量抗原淋巴結(jié)內(nèi)注射免疫家兔·一般選擇～–先在其兩腳墊注射完全福氏佐劑(卡介苗每兔合5mg)；–10天后，見(jiàn)胭窩淋巴結(jié)腫大，然后將抗原注射至腫大的淋巴結(jié)內(nèi)，***次注射抗原用完全福氏佐劑混合乳化；–以后每隔20天用不完全福氏佐劑乳化抗原，以同樣方法加強(qiáng)2次，各次注射的抗原均為25～50?g；–末次注射兩周后兔耳放血測(cè)價(jià)(可達(dá)1:128)。舉例3：豚鼠和大鼠的免疫·初次免疫–用10～100?gAg加入FCA，在背部皮內(nèi)注射4～6點(diǎn)，每點(diǎn)，也可肌肉內(nèi)或皮下注射；·加強(qiáng)免疫–每隔7～8天，將10～50?gAg于PBS或FIA中。在肌肉、皮下或靜脈注射；·抗體效價(jià)的檢測(cè)(4)免疫劑量·抗原性強(qiáng)的抗原量應(yīng)小，過(guò)大反會(huì)引起免疫***；·免疫周期長(zhǎng)的動(dòng)物可少量多次注射，短的可大量少次。(4)免疫劑量(續(xù))·各種動(dòng)物***免疫抗原劑量和加強(qiáng)免疫的劑量(5)抗體效價(jià)的檢測(cè)多采免疫雙向擴(kuò)散法【基本原理】·指抗原和抗體在同一凝膠內(nèi)都擴(kuò)散，彼此相遇后形成特異性的沉淀線。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="天津多色熒光免疫組化分析服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748330079/709907.png"></div><p> 80年代到90年代相繼又有新的熒光素出現(xiàn)如R-藻紅朊、B-藻紅朊、C-藻青蛋白、cy2、cy3、cy5和cy7等均在流式細(xì)胞儀和激光共聚焦顯微鏡中廣泛應(yīng)用。由于免疫熒光組織化學(xué)的特異性，快速性和在細(xì)胞水平定位的準(zhǔn)確性，已在免疫學(xué)、微生物學(xué)、病理學(xué)、**學(xué)以及臨床檢驗(yàn)等許多方面得到廣泛應(yīng)用，日益發(fā)揮重要作用。[1]免疫組織化學(xué)技術(shù)現(xiàn)狀與展望編輯免疫熒光組織化學(xué)技術(shù)經(jīng)過(guò)半個(gè)多世紀(jì)的不斷改進(jìn)和創(chuàng)新，已成為現(xiàn)代研究生物和醫(yī)學(xué)的重要手段之一。由于免疫熒光技術(shù)與形態(tài)、機(jī)能相結(jié)合不斷完善和發(fā)展，尤其是合成了多種新熒光素與抗體容易結(jié)合，且結(jié)合物穩(wěn)定?？梢院虵ITC結(jié)合進(jìn)行免疫熒光組織化學(xué)雙標(biāo)記或三標(biāo)記。至今，它已和親合化學(xué)技術(shù)如SPA、Biotin以及avidin、ConA相結(jié)合，應(yīng)用領(lǐng)域也日益擴(kuò)大，又與現(xiàn)代的電子計(jì)算機(jī)、掃描電鏡技術(shù)、共聚焦顯微鏡、熒光***細(xì)胞分類(lèi)器（FACS）以及數(shù)碼相機(jī)攝影技術(shù)的應(yīng)用，使得快速性、簡(jiǎn)便性有了更大的提高，使得定量更加準(zhǔn)確。90年代又開(kāi)展了熒光原位末端標(biāo)記和熒光原位雜交技術(shù)，使得應(yīng)用范圍更廣。雖然免疫組織化學(xué)技術(shù)有了很大的發(fā)展，如免疫金銀法、免疫膠體金法、SP、Evision二步法和CSA法，但由于它有自己獨(dú)特的優(yōu)點(diǎn)：特異性強(qiáng)。江蘇七色免疫組化分析服務(wù)</p><p style="text-indent:25px">天津多色熒光免疫組化分析服務(wù)</p><p> 由于技術(shù)上的限制，只有直接法、間接法等敏感性不高的技術(shù)，那時(shí)的抗體只能稀釋幾倍、幾十倍；現(xiàn)在由于ABC法或SP三步法的出現(xiàn)，使抗體稀釋上千倍、上萬(wàn)倍甚至上億倍仍可在組織細(xì)胞中與抗原結(jié)合，這樣高敏感性的抗體抗原反應(yīng)，使免疫組化方法越來(lái)越方便地應(yīng)用于常規(guī)病理診斷工作。3）定位準(zhǔn)確、形態(tài)與功能相結(jié)合該技術(shù)通過(guò)抗原抗體反應(yīng)及呈色反應(yīng)，可在組織和細(xì)胞中進(jìn)行抗原的準(zhǔn)確定位，因而可同時(shí)對(duì)不同抗原在同一組織或細(xì)胞中進(jìn)行定位觀察，這樣就可以進(jìn)行形態(tài)與功能相結(jié)合的研究，對(duì)病理學(xué)領(lǐng)域開(kāi)展深入研究是十分有意義的。7、從蛋白水平檢測(cè)角度，免疫組化技術(shù)與Westernblotting、ELISA的異同1）Westernblotting蛋白質(zhì)免疫印跡，也是利用抗體抗原反應(yīng)原理，結(jié)合化學(xué)發(fā)光等技術(shù)來(lái)檢查組織或細(xì)胞樣品內(nèi)蛋白含量的檢測(cè)方法。與免疫組化技術(shù)相比，定量可能更加準(zhǔn)確；當(dāng)然Westernblotting也可定性和定位（通過(guò)提取膜蛋白或**白、胞漿蛋白分別檢測(cè)其中抗原含量，進(jìn)而間接反映它們的定位），但敏感性遠(yuǎn)遠(yuǎn)低于免疫組化技術(shù)。2）ELISA酶聯(lián)免疫吸附試驗(yàn)，也是利用抗體-抗原-抗原結(jié)合反應(yīng)原理來(lái)檢查體液或組織勻漿中蛋白含量的檢測(cè)。與免疫組化技術(shù)相比，定量**準(zhǔn)確。天津多色熒光免疫組化分析服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司位于上海市，創(chuàng)立于2003-01-22。上海朝瑞科技致力于為客戶提供質(zhì)量的[ "科研技術(shù)服務(wù)", 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