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href="http://www.cdcfah.com/cp/third-nongye_nlmyxmhz/">農(nóng)林牧漁項(xiàng)目合作</a>?江蘇多標(biāo)記免疫組化技術(shù)服務(wù) **** 上海朝瑞生物科技供應(yīng)</p><div id="v8sjfwi" class="leftbox clearfix"><h1>江蘇多標(biāo)記免疫組化技術(shù)服務(wù) **** 上海朝瑞生物科技供應(yīng)</h1><div id="cqhyfgx" class="proBig"><div id="gjqsj3b" class="bigimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748330079/709907.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div><div id="k3d3y9b" class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748330079/709907.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="xzypqm8" 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>江蘇多標(biāo)記免疫組化技術(shù)服務(wù),免疫組化</p><p><span>***更新：</span>2020-08-14 22:13:29</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="s3gua72" 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="hzwy74x" class="page-proshow02"><p class="title">詳細(xì)說(shuō)明</p><div id="leliels" class="box"><p><p> 在堿性條件下，易與蛋白質(zhì)的賴氨酸?-氨基反應(yīng)而標(biāo)記在蛋白分子上。(4)碘化丙啶(PI)·是常用的DNA熒光標(biāo)記探針，可作為FITC的胞核對(duì)比染色?！I可嵌入到雙鏈DNA和RNA堿基對(duì)中并與之結(jié)合，但對(duì)堿基無(wú)特異性選擇?！け容^大吸收光譜是493nm，比較大發(fā)射光譜是630nm，呈紅色熒光，江蘇多標(biāo)記免疫組化技術(shù)服務(wù)。2、熒光抗體的保存·一要防止抗體失活，江蘇多標(biāo)記免疫組化技術(shù)服務(wù)，二要保持熒光素不脫落和不受激發(fā)猝滅。–一般認(rèn)為0～4?C可保存1～2年，-20?C可保存3～4年。–要小量分裝，防止反復(fù)凍融。–保存前需加防腐劑(濃度為1:5000～10000的硫柳汞或1:1000～5000疊氮化鈉)。3、免疫熒光的染色方法·免疫熒光染色法常用的有–直接法–間接法?原理：將熒光素標(biāo)記在相應(yīng)的抗體上，直接與相應(yīng)抗原反應(yīng)(用來(lái)檢測(cè)未知抗原)，江蘇多標(biāo)記免疫組化技術(shù)服務(wù)。?直接免疫熒光法的操作步驟·標(biāo)本的處理：–細(xì)胞涂片、細(xì)胞爬片浸入冷**或4%的多聚甲醛固定10min，然后用PBST(含pH)漂洗5min×3/次；–石蠟切片經(jīng)脫蠟、梯度酒精脫水后，進(jìn)行抗原修復(fù)，然后用PBST漂洗5min×3/次；·2%BSA或10%BSA37?C濕盒內(nèi)封閉30min·抗體染色：–在標(biāo)本片上滴加適當(dāng)稀釋的熒光標(biāo)記抗體(1:8或1:16稀釋)，放在濕盒中，37?C孵育30min；?直接免疫熒光法的操作步驟(續(xù))·PBS。江蘇多標(biāo)記免疫組化技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇多標(biāo)記免疫組化技術(shù)服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748330079/709907.png"></div><p> ）問(wèn)題及其解答：如何降低非特異性熒光染色和避免陰性著色？1、非特異性染色產(chǎn)生原因及其解決方案：⑴游離熒光素殘留在二抗中。一部分熒光素未與蛋白質(zhì)結(jié)合，形成了聚合物和衍化物，而不能被透析除去。二抗配置時(shí)用透析法或?qū)游龇ǚ蛛x熒光素標(biāo)記的二抗和游離的二抗。購(gòu)買(mǎi)高質(zhì)量、高純度的熒光素二抗。⑵抗體以外的血清蛋白與熒光素結(jié)合形成熒光素脲蛋白，可與組織成分結(jié)合。⑶組織抗原封閉不全。除去檢查的抗原以外，組織中還可能存在類(lèi)屬抗原（如Forssman氏抗原），可與組織中特異性抗原以外之之相應(yīng)抗體結(jié)合。延長(zhǎng)血清封閉時(shí)間。⑷從組織中難于提純抗原性物質(zhì)，所以制備的免疫血清中往往混雜一些抗其他組織成分的抗體，以致容易混淆。⑸抗體分子上標(biāo)記的熒光素分子太多，這種過(guò)量標(biāo)記的抗體分子帶過(guò)多的陰離子，可吸附于正常組織上而呈現(xiàn)非特異性染色。⑹熒光素不純、標(biāo)本固定不當(dāng)?shù)?。⑺一抗和二抗孵育條件和濃度不合適。調(diào)整一抗和二抗孵育條件和濃度至比較好。⑻一抗和二抗孵育后的清洗不充分。增加清洗次數(shù)和延長(zhǎng)清洗時(shí)間。2、陰性染色產(chǎn)生的原因有：⑴一抗和二抗?jié)舛炔缓线m或孵育條件不妥。⑵熒光素提前衰退。江蘇多標(biāo)記免疫組化技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇多標(biāo)記免疫組化技術(shù)服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750849610/4353685.png"></div><p> 問(wèn)題及其解答：產(chǎn)生組織切片非特異性染色的原因有哪些？如何解決？1、抗體孵育時(shí)間過(guò)長(zhǎng)、抗體濃度高易增加背景著色。這可通過(guò)縮短一抗/二抗孵育時(shí)間、稀釋抗體來(lái)控制。這是**重要的一條。2、一抗用多克隆抗體易出現(xiàn)非特異性著色，建議試用單克隆抗體看看。3、內(nèi)源性過(guò)氧化物酶和生物素在肝臟、腎臟等組織含量很高（含血細(xì)胞多的組織），需要通過(guò)延長(zhǎng)滅活時(shí)間和增加滅活劑濃度來(lái)降低背景染色；4、非特異性組分與抗體結(jié)合，這需要通過(guò)延長(zhǎng)二抗來(lái)源的動(dòng)物免疫血清封閉時(shí)間和適當(dāng)增加濃度來(lái)加強(qiáng)封閉效果；5、DAB孵育時(shí)間過(guò)長(zhǎng)或濃度過(guò)高；6、PBS沖洗不充分，殘留抗體結(jié)果增強(qiáng)著色，在一抗/二抗/SP孵育后的浸洗尤為重要；7、標(biāo)本染色過(guò)程中經(jīng)常出現(xiàn)干片，這容易增強(qiáng)非特異性著色。我的實(shí)際解決方案：以上的分析可能對(duì)于初學(xué)者還是不容易的，下面我就把我的排除實(shí)驗(yàn)的具體過(guò)程與大家進(jìn)行分享、交流和討論。1、首先排除抗體孵育條件。一般來(lái)說(shuō)，著色效果的好壞與抗體的孵育條件是密不可分的，由于用SP法已經(jīng)做出來(lái)過(guò)一次且效果不錯(cuò)，因此對(duì)于同樣組織的同一抗原進(jìn)行分析，這種因素可以先排除。2、其次，DAB孵育時(shí)間是否太長(zhǎng)？做出來(lái)那一次我是孵育8min，后來(lái)也是8-10min。</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="江蘇多標(biāo)記免疫組化技術(shù)服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748427735/7999973.png"></div><p style="text-indent:25px">TSA技術(shù)(Tyramide Signal Amplification?，酪胺信號(hào)放大技術(shù))是做多重免疫熒光染色的一種多快好省的方法，它使同一張片子上所有的靶標(biāo)都可以選用特異性高的兔單克隆抗體，搭配同一支抗兔的HRP二抗就可以進(jìn)行實(shí)驗(yàn)，而且信號(hào)放大的倍數(shù)**增強(qiáng)。如果想要運(yùn)用這個(gè)技術(shù)，需要先做四個(gè)方面的準(zhǔn)備工作：嚴(yán)格驗(yàn)證的高特異性抗體：所有CST的IHC-P驗(yàn)證過(guò)的抗體都滿足mIHC(多重?zé)晒饷庖呓M化)高特異性的要求;HRP偶聯(lián)的針對(duì)一抗種屬亞型特異的二抗;酪胺熒光素(Perkin Elmer,Thermo Fisher Scientific等供應(yīng)商);多光譜成像系統(tǒng)(PerkinElmer等)。江蘇多標(biāo)記免疫組化技術(shù)服務(wù)</p><p style="text-indent:25px">江蘇多標(biāo)記免疫組化技術(shù)服務(wù)</p><p> –新載玻片上有油污，要用洗液浸泡12～24h，自來(lái)水沖洗后再用蒸餾水清洗；用綢布擦干或烤箱烤干。清潔的載玻片再用粘附劑處理?！こＳ玫恼掣絼┯校酣CAPES（3-氨丙基三乙氧基硅烷）–Polv-L-Lysine（多聚賴氨酸）–鉻明膠溶液APES(3-aminopropyltriethoxysilane)3-氨丙基三乙氧基硅烷·現(xiàn)用現(xiàn)配。用純**或甲醇配制2%APES(v/v);·將洗凈的玻片浸于此液中20～30秒鐘；·取出稍停片刻，再入純**酮溶液洗去未結(jié)合的APES，置通風(fēng)櫥中晾干或60?C烤箱烤干。Polv-L-Lysine(多聚賴氨酸)·將清潔玻片浸于100?g/ml(10%w/v)的多聚賴氨酸溶液中(去離子水稀釋)，37?C放置30min，然后60?C烤箱烘烤1h或室溫過(guò)夜干燥。裝盒備用。鉻明膠溶液·試劑：鉻明礬(chromealum)明膠(gelatine)蒸餾水500ml·先將鉻明礬溶解于40?C少量蒸餾水中，再加入明膠及蒸餾水，可在70?C水浴中使明膠溶化，攪拌均勻后，即可使用。如有殘?jiān)蛇^(guò)濾后再用。·用時(shí)稍加溶化，切片浸入2～3min，過(guò)夜晾干。免疫細(xì)胞化學(xué)技術(shù)冰凍切片·冰凍切片是指將組織在冷凍狀態(tài)下直接切片。·在切片前組織不經(jīng)過(guò)任何化學(xué)藥品處理或加熱過(guò)程。–縮短了制片時(shí)間–抗原性不受損失·對(duì)穩(wěn)定性差的抗原。江蘇多標(biāo)記免疫組化技術(shù)服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司創(chuàng)建于2003-01-22，注冊(cè)資金 700-1000萬(wàn)元，是一家專(zhuān)注1.發(fā)布科研技術(shù) 2.發(fā)布應(yīng)用技術(shù) 3.發(fā)布專(zhuān)業(yè)專(zhuān)長(zhǎng) 4.發(fā)布科研成果 5.發(fā)布我的需求 6.發(fā)布篩選檢驗(yàn)檢測(cè)服務(wù) 7.發(fā)布協(xié)同代研發(fā)服務(wù) 8.發(fā)布升級(jí)改造產(chǎn)品服務(wù) 9.發(fā)布實(shí)驗(yàn)室及儀器設(shè)備共享 10.發(fā)布培訓(xùn)基地共享 11.發(fā)布工廠代加工及共享車(chē)間 12.發(fā)布科研/項(xiàng)目團(tuán)隊(duì)招聘13.發(fā)布研究生婚戀 14.發(fā)布項(xiàng)目整包服務(wù) 15.發(fā)布資源交換業(yè)務(wù)的公司。公司目前擁有5~10人員工，為員工提供廣闊的發(fā)展平臺(tái)與成長(zhǎng)空間，為客戶提供高質(zhì)高效的產(chǎn)品服務(wù)，深受員工與客戶好評(píng)。誠(chéng)實(shí)、守信是對(duì)企業(yè)的經(jīng)營(yíng)要求，也是我們做人的基本準(zhǔn)則。公司致力于打造***的[ "科研技術(shù)服務(wù)", "應(yīng)用技術(shù)服務(wù)", "科研成果轉(zhuǎn)化", "科學(xué)產(chǎn)品銷(xiāo)售" ]。公司力求給客戶提供***質(zhì)量服務(wù)，我們相信誠(chéng)實(shí)正直、開(kāi)拓進(jìn)取地為公司發(fā)展做正確的事情，將為公司和個(gè)人帶來(lái)共同的利益和進(jìn)步。經(jīng)過(guò)幾年的發(fā)展，已成為[ "科研技術(shù)服務(wù)", "應(yīng)用技術(shù)服務(wù)", "科研成果轉(zhuǎn)化", "科學(xué)產(chǎn)品銷(xiāo)售" ]行業(yè)**企業(yè)。</p></p><br /><p>文章來(lái)源地址: http://www.cdcfah.com/cp/917779.html</p></div></div></div></div></div><div id="qyk2vct" class="page-right twoColRt"><div id="yxjqc7u" class="box01 margin10 clearfix"><h3><a href="http://www.cdcfah.com/cp/">猜你喜歡</a></h3><div id="ga2p2xr" 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