江蘇磷酸化蛋白Western Blot免疫印跡服務(wù) 來電咨詢上海朝瑞生物科技供應(yīng)-b2b網(wǎng)站_b2b電子商務(wù)平臺(tái)【買產(chǎn)品】-上?？谄蚴称酚邢薰?/title><meta name="description" content="免疫印跡法（immunoblotting）因與Southern早先建立的檢測核酸的印跡方法Southernblot相類似，江蘇磷酸化蛋白WesternBlot免疫印跡服務(wù)，亦被稱為Westernblot。免疫印跡（westernblotti"><meta name="keywords" content="江蘇磷酸化蛋白Western Blot免疫印跡服務(wù),western blot"><link rel="canonical" href="http://www.cdcfah.com/"/><meta name="applicable-device" content="pc"><link rel="shortcut icon" href="/static/new_net_01/images/favicon.ico"><link rel="stylesheet" href="/static/new_net_01/css/slick.css" /><link rel="stylesheet" href="/static/new_net_01/css/addnew.css"><link rel="stylesheet" href="/static/new_net_01/css/sidenav.css"><link rel="stylesheet" href="/static/new_net_01/css/style.css" /><link rel="stylesheet" href="/static/new_net_01/css/tab.css"><link rel="stylesheet" href="/static/new_net_01/css/404.css"> <meta name="baidu-site-verification" content="OBVzrC9aaQ" /> <meta http-equiv="Cache-Control" 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Blot免疫印跡服務(wù),western blot</p><p><span>***更新：</span>2020-07-31 22:13:35</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="4iqgwc2" 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="4404o2q" class="page-proshow02"><p class="title">詳細(xì)說明</p><div id="uqo4oui" class="box"><p><p> 免疫印跡法（immunoblotting ）因與Southern早先建立的檢測核酸的印跡方法Southern </p> <p> blot相類似，江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)，亦被稱為Western blot。免疫印跡（western blotting）是在蛋白質(zhì)電泳分離和抗原抗體檢測的基礎(chǔ)上發(fā)展起來一項(xiàng)檢測蛋白質(zhì)的技術(shù)。它將SDS聚丙烯酰胺凝膠電泳的辨力與抗原抗體反應(yīng)的特異性相結(jié)合。典型的印跡實(shí)驗(yàn)包括三個(gè)步驟①蛋白質(zhì)的電泳分離；②將電泳后凝膠上的蛋白質(zhì)轉(zhuǎn)移至固體膜上，用非特異性，江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)，非反應(yīng)活性分子封閉固體膜上未吸附蛋白質(zhì)區(qū)域；③免疫學(xué)檢測，江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)。免疫印跡克服了聚丙烯酰胺凝膠電泳后直接在凝膠上進(jìn)行免疫學(xué)分析的弊端，極大地提高了其利用率、分辨率和靈敏度，使其成為使用**的免疫化學(xué)方法之一。江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇磷酸化蛋白Western Blot免疫印跡服務(wù),western blot" src="http://tyunfile.71360.com/UploadFile/shchaorui/637115400031748047/9643065.png"></div><p> 全自動(dòng)western blot無需凝膠、無需轉(zhuǎn)印，放入您的樣品，按個(gè)按鈕，然后就沒事了。首先，將樣品與Simple </p> <p> Western樣品緩沖液混合，至終濃度1 μg/μL，還原并變性，然后將制備好的樣品、一抗和二抗、化學(xué)發(fā)光底物分配到384孔分析板的指定孔中。之后，將Simple </p> <p> Western分析緩沖液、毛細(xì)管和制備好的分析板放入Simon內(nèi)，它自動(dòng)開展所有分析步驟。當(dāng)?shù)鞍籽刂?xì)管內(nèi)一個(gè)堆積和分離基質(zhì)遷移時(shí)，它們分離。分離后的蛋白固定到毛細(xì)管壁上。之后利用一抗鑒定目標(biāo)蛋白，用HRP結(jié)合的二抗和化學(xué)發(fā)光底物檢測蛋白。系統(tǒng)自動(dòng)報(bào)告檢測蛋白的分子量和信號(hào)。每次實(shí)驗(yàn)可同時(shí)分析**多12個(gè)樣品，結(jié)果可在3-5小時(shí)內(nèi)獲得。15-150 kDa的目標(biāo)蛋白均可檢測。軟件報(bào)告每個(gè)檢測蛋白的分子量、面積、面積百分比和信噪比。對于傳統(tǒng)的Western blot而言，結(jié)果的重復(fù)性一直是個(gè)挑戰(zhàn)，因?yàn)槿狈?biāo)準(zhǔn)化的操作，且多個(gè)操作步驟引入了實(shí)驗(yàn)可變性。而Simple Western分析是完全自動(dòng)化的，因此結(jié)果的重復(fù)性更好。整體的定量也**改善了，因?yàn)槭∪チ宿D(zhuǎn)印步驟，從而避免了蛋白轉(zhuǎn)移的不一致。上海熒光Western Blot檢測分析服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇磷酸化蛋白Western Blot免疫印跡服務(wù),western blot" src="http://tyunfile.71360.com/UploadFile/shchaorui/637113662702265625/5703187.jpg"></div><p> 轉(zhuǎn)膜(Transfer) </p> <p> 推薦在Western實(shí)驗(yàn)中選用PVDF膜。硝酸纖維素膜(NC膜)也可以使用，但硝酸纖維素膜比較脆，在操作過程中特別是用鑷子夾取等過程中容易裂開。膜的使用請參考生產(chǎn)商的推薦使用步驟。 </p> <p> 通常如果使用Bio-Rad的標(biāo)準(zhǔn)濕式轉(zhuǎn)膜裝置，可以設(shè)定轉(zhuǎn)膜電流為300-400mA，轉(zhuǎn)膜時(shí)間為30-60分鐘。也可以在15-20mA轉(zhuǎn)膜過夜。具體的轉(zhuǎn)膜時(shí)間要根據(jù)目的蛋白的大小而定，目的蛋白的分子量越大，需要的轉(zhuǎn)膜時(shí)間越長，目的蛋白的分子量越小，需要的轉(zhuǎn)膜時(shí)間越短。在轉(zhuǎn)膜過程中，特別是高電流快速轉(zhuǎn)膜時(shí)，通常會(huì)有非常嚴(yán)重的發(fā)熱現(xiàn)象，比較好把轉(zhuǎn)膜槽放置在冰浴中進(jìn)行轉(zhuǎn)膜。 </p> <p> 轉(zhuǎn)膜的效果可以觀察所使用的預(yù)染蛋白質(zhì)分子量標(biāo)準(zhǔn)，通常分子量比較大的1-2條帶較難全部轉(zhuǎn)到膜上。轉(zhuǎn)膜的效果也可以用麗春紅染色液(P0022)對膜進(jìn)行染色，以觀察實(shí)際的轉(zhuǎn)膜效果。也可以用考馬斯亮藍(lán)快速染色液(P0017)對完成轉(zhuǎn)膜的SDS-PAGE膠進(jìn)行染色，以觀察蛋白的殘留情況。 </p><p> 1.為SDS—聚丙烯酰胺凝膠電泳（SDS-PAGE）?？乖鹊鞍讟悠方?jīng)SDS處理后帶陰電荷，在聚丙烯酰膪凝膠中從陰極向陽極泳動(dòng)，分子量越小，泳動(dòng)速度就越快。此階段分離效果肉眼不可見（只有在染色后才顯出電泳區(qū)帶）。2.電轉(zhuǎn)移。將在凝膠中已經(jīng)分離的條帶轉(zhuǎn)移至硝酸纖維素膜上，選用低電壓（100V）和大電流（1～2A），通電45min轉(zhuǎn)移即可完成。此階段分離的蛋白質(zhì)條帶肉眼仍不可見。3.酶免疫定位。將印有蛋白質(zhì)條帶的硝酸纖維素膜（相當(dāng)于包被了抗原的固相載體）依次與特異性抗體和酶標(biāo)第二抗體作用后，加入能形成不溶性顯色物的酶反應(yīng)底物，使區(qū)帶染色。常用的HRP底物為3，3，—二氨基聯(lián)苯胺（呈棕色）和4-氯-1-萘酚（呈藍(lán)紫色）。陽性反應(yīng)的條帶清晰可辨，并可根據(jù)SDS-PAGE時(shí)加人的分子量標(biāo)準(zhǔn)，確定各組分的分子量。本法綜合了SDS-PAGE的辨力和ELISA法的高特異性和敏感性，是一個(gè)有效的分析手段，不僅廣泛應(yīng)用于分析抗原組分及其免疫活性，并可用于疾病的診斷。在**中此法作為確診試驗(yàn)?？乖?jīng)電泳轉(zhuǎn)移在硝酸纖維素膜上后，將膜切成小條，配合酶標(biāo)抗體及顯色底物制成的試劑盒可方便地在實(shí)驗(yàn)室中供檢測用。 </p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="江蘇磷酸化蛋白Western Blot免疫印跡服務(wù),western blot" src="http://tyunfile.71360.com/UploadFile/shchaorui/637115400030400391/2584407.png"></div><p> 轉(zhuǎn)膜完畢后，立即把蛋白膜放置到預(yù)先準(zhǔn)備好的Western洗滌液(P0023C)中，漂洗1-2分鐘，以洗去膜上的轉(zhuǎn)膜液。從轉(zhuǎn)膜完畢后所有的步驟，一定要注意膜的保濕，避免膜的干燥，否則極易產(chǎn)生較高的背景。用微型臺(tái)式真空泵或滴管等吸盡封閉液，立即加入稀釋好的一抗，室溫或4℃在側(cè)擺搖床上緩慢搖動(dòng)孵育一小時(shí)。如果一抗孵育一小時(shí)效果不佳，可以4℃緩慢搖動(dòng)孵育過夜?；蚋鼡?jù)抗體的說明選擇適當(dāng)?shù)姆跤郎囟群蜁r(shí)間。微型臺(tái)式真空泵或滴管等吸盡洗滌液，立即加入稀釋好的二抗，室溫或4℃在側(cè)擺搖床上緩慢搖動(dòng)孵育一小時(shí)。回收二抗。加入Western洗滌液，在側(cè)擺搖床上緩慢搖動(dòng)洗滌5-10分鐘。吸盡洗滌液后，再加入洗滌液洗滌5-10分鐘。共洗滌3次。如果結(jié)果背景較高可以適當(dāng)延長洗滌時(shí)間并增加洗滌次數(shù)。使用 </p> <p> ECL化學(xué)發(fā)光試劑來檢測蛋白。壓片可以采用**的壓片暗盒進(jìn)行。洗片時(shí)可以使用X光片自動(dòng)洗片機(jī)。如果沒有自動(dòng)洗片機(jī)，可以用顯影定影試劑盒自行配制顯影液和定影液進(jìn)行手工洗片。X光片推薦選用柯達(dá)原裝的生物實(shí)驗(yàn)**柯達(dá)X-OMAT BT膠片。上海長條帶Western Blot免疫印跡服務(wù)</p><p style="text-indent:25px">江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)</p><p> 收集蛋白樣品(Protein sample preparation) </p> <p> 可以使用適當(dāng)?shù)牧呀庖?，Western及IP細(xì)胞裂解液、RIPA裂解液等，裂解貼壁細(xì)胞、懸浮細(xì)胞或組織樣品。對于某些特定的亞細(xì)胞組份蛋白，例如細(xì)胞**白、細(xì)胞漿蛋白、線粒體蛋白等，可以參考相關(guān)文獻(xiàn)提取這些亞細(xì)胞組份蛋白，也可以使用試劑盒進(jìn)行抽提。收集完蛋白樣品后，為確保每個(gè)蛋白樣品的上樣量一致，需要測定每個(gè)蛋白樣品的蛋白濃度。根據(jù)所使用的裂解液的不同，需要采用適當(dāng)?shù)牡鞍诐舛葴y定方法。因?yàn)椴煌牡鞍诐舛葴y定方法對于一些去垢劑和還原劑等的兼容性差別很大。如果使用Western及IP細(xì)胞裂解液或RIPA裂解液，可以使用BCA蛋白濃度測定試劑盒江蘇磷酸化蛋白Western Blot免疫印跡服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司坐落在上海市松江區(qū)廣富林路697號(hào)昂立大廈2113室，是一家專業(yè)的1.發(fā)布科研技術(shù) 2.發(fā)布應(yīng)用技術(shù) 3.發(fā)布專業(yè)專長 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)公司。目前我公司在職員工達(dá)到5~10人人，是一個(gè)有活力有能力有創(chuàng)新精神的高效團(tuán)隊(duì)。誠實(shí)、守信是對企業(yè)的經(jīng)營要求，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)品銷售" ]。公司力求給客戶提供***質(zhì)量服務(wù)，我們相信誠實(shí)正直、開拓進(jìn)取地為公司發(fā)展做正確的事情，將為公司和個(gè)人帶來共同的利益和進(jìn)步。經(jīng)過幾年的發(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)品銷售" ]行業(yè)**企業(yè)。</p></p><br /><p>文章來源地址: http://www.cdcfah.com/cp/767202.html</p></div></div></div></div></div><div id="g2m2goc" class="page-right twoColRt"><div id="qeukyw2" class="box01 margin10 clearfix"><h3><a href="http://www.cdcfah.com/cp/">猜你喜歡</a></h3><div id="qec2y4o" class="recomPic likePro"><a href="http://www.cdcfah.com/cp/814588.html"><div><img src="http://tyunfile.71360.com/UpLoadFile/2019/11/1/14/637082168864691056_lgpj_5374157.jpg" alt="福建ZF配件哪家強(qiáng) 信息推薦自業(yè)物資供應(yīng)" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></div><p>福建ZF配件哪家強(qiáng) 信息推薦自業(yè)物資供應(yīng)</p></a></div><div id="a2022qg" class="recomPic likePro"><a href="http://www.cdcfah.com/cp/846569.html"><div><img 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