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src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750107422/8042334.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div><div id="qe2kp7l" class="smallimg"><div><p class="pic"><img src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750107422/8042334.png" class="vcenter" onerror="javascript:this.src='/static/new_net_01/images/nopic.jpg';"></p></div></div></div><div id="dczw3s3" 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>上海多靶點(diǎn)免疫組化染色服務(wù),免疫組化</p><p><span>***更新：</span>2020-08-17 03:11:40</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="weqx8sr" 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="ztqnprn" class="page-proshow02"><p class="title">詳細(xì)說(shuō)明</p><div id="ys3oyf2" class="box"><p><p> 免疫細(xì)胞化學(xué)技術(shù)組織印片·將潔凈載玻片輕壓于已暴露病灶的新鮮組織切面，細(xì)胞即粘附于玻片，晾干后浸入冷**或醋酸-乙醇固定10min，自然干燥后染片或-20℃保存。免疫細(xì)胞化學(xué)技術(shù)細(xì)胞培養(yǎng)片(細(xì)胞爬片)·貼壁細(xì)胞培養(yǎng)時(shí)，置蓋片于培養(yǎng)瓶中，使細(xì)胞在蓋片上生長(zhǎng)，達(dá)適當(dāng)密度后取出固定(**-20?C固定10～20min)，再進(jìn)行免疫染色。–蓋片的處理方法同載玻片的處理，但泡酸時(shí)間2h即可。–為了防止細(xì)胞脫片，可用多聚賴氨酸處理。免疫細(xì)胞化學(xué)技術(shù)細(xì)胞涂片·大多數(shù)細(xì)胞涂片由細(xì)胞懸液制成，包括：–血液、尿液、腦脊液；–體腔積液；–組織穿刺吸取，如骨髓、淋巴結(jié)或其他實(shí)質(zhì)性組織–懸浮培養(yǎng)的細(xì)胞或貼壁細(xì)胞經(jīng)消化后形成的懸液。免疫細(xì)胞化學(xué)技術(shù)細(xì)胞涂片(續(xù))·細(xì)胞涂片的方法：–手涂法·將細(xì)胞濃度調(diào)節(jié)到106/ml左右，可直接涂于載玻片上，但要均勻、不重疊，上海多靶點(diǎn)免疫組化染色服務(wù)?！D片范圍應(yīng)小于1cm直徑，以節(jié)約試劑。–涂片機(jī)涂片法·將細(xì)胞樣品制成2×105～6/ml細(xì)胞懸液，上海多靶點(diǎn)免疫組化染色服務(wù)，吸取50～100?l(1～2×104～5cells)加入涂片機(jī)內(nèi)，上海多靶點(diǎn)免疫組化染色服務(wù)，1000rpm離心2min后細(xì)胞就均勻分布于玻片上。免疫細(xì)胞化學(xué)技術(shù)常用染色方法根據(jù)標(biāo)記物的不同分為免疫熒光法、免疫酶標(biāo)法、親和組織化學(xué)法。上海多靶點(diǎn)免疫組化染色服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海多靶點(diǎn)免疫組化染色服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750107422/8042334.png"></div><p> 避免了共價(jià)連接對(duì)抗體和酶活性的損害，提高了方法的敏感性，而且節(jié)省一抗的用量。但抗酶抗體不易純化。幾點(diǎn)說(shuō)明·一抗(假設(shè)來(lái)自種屬A)的稀釋度可大些，使抗體的兩個(gè)Fab段均與組織抗原結(jié)合?！ざ埂獦蚩贵w(抗種屬A的IgG抗體)應(yīng)過(guò)量，使其Fab段一個(gè)與一抗結(jié)合，另一個(gè)則游離?！ひ蚩姑缚贵w與一抗均系種屬AIgG，具有相同的抗原性，所以橋抗體游離的Fab能與抗酶抗體結(jié)合，起橋作用，將其連接在與組織抗原結(jié)合的一抗上。(2)非標(biāo)記抗體酶法——PAP法·與酶橋法相似，不同的是，PAP法將酶橋法的第3、4步并為1步，用PAP復(fù)合物代替;·PAP是離體制備的復(fù)合物(HRP--抗HRP)?！わ@色與酶橋法相同，PAP復(fù)合物中的過(guò)氧化物酶催化底物水解，形成不溶性終產(chǎn)物。PAP法評(píng)價(jià)·PAP法比直接法、間接法、酶橋法更敏感。特別適用于石蠟切片中微量抗原和抗原性減弱抗原的檢測(cè)。–缺點(diǎn)：步驟較多，時(shí)間較長(zhǎng)，不適用于臨床常規(guī)檢查。·由于常做石蠟切片，故可用于回顧性研究。免疫細(xì)胞化學(xué)技術(shù)親和組織化學(xué)法·是以一種物質(zhì)對(duì)某種組織成分具有高度親合力為基礎(chǔ)?！み@種方法敏感性更高，有利于微量抗原(抗體)在細(xì)胞或亞細(xì)胞水平的定位。生物素—抗生物素染色法【原理】·生物素(biotin)又稱維生素H。上海多靶點(diǎn)免疫組化染色服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海多靶點(diǎn)免疫組化染色服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899750458985/5221034.png"></div><p> 這方面我有慘痛教訓(xùn)，當(dāng)時(shí)我買的抗體稀釋液偏酸，結(jié)果背景一片黃（未見(jiàn)特異性染色），建議pH在。（中性及弱堿性條件（pH7-8）有利于免疫復(fù)合物的形成，而酸性條件則有利于分解；低離子強(qiáng)度有利于免疫復(fù)合物的形成，而高離子強(qiáng)度則有利于分解）9）拍照有條件的話比較好立即拍照，若不能及時(shí)拍照，也要封好片和用指甲油封固，保持避光和濕度。使用熒光顯微鏡注意嚴(yán)格按照熒光顯微鏡出廠說(shuō)明書要求進(jìn)行操作，不要隨意改變程序；應(yīng)在暗室中進(jìn)行檢查；防止紫外線對(duì)眼睛的損害，在調(diào)整光源時(shí)應(yīng)戴上防護(hù)眼鏡；檢查時(shí)間每次以1~2h為宜，超過(guò)90min，超高壓汞燈發(fā)光強(qiáng)度逐漸下降，熒光減弱；標(biāo)本受紫外線照射3~5min后，熒光也明顯減弱或褪色；激發(fā)光長(zhǎng)時(shí)間的照射，會(huì)發(fā)生熒光的衰減和淬滅現(xiàn)象；所以**多不得超過(guò)2~3h；熒光顯微鏡光源壽命有限，標(biāo)本應(yīng)集中檢查，以節(jié)省時(shí)間，保護(hù)光源。天熱時(shí)，應(yīng)加電扇散熱降溫，新?lián)Q燈泡應(yīng)從開始就記錄使用時(shí)間。燈熄滅后欲再啟用時(shí)，須待燈光充分冷卻后才能點(diǎn)燃。***中應(yīng)避免數(shù)次點(diǎn)燃光源。關(guān)閉汞燈至少在開啟15-30分鐘后；標(biāo)本染色后立即觀察，因時(shí)間久了熒光會(huì)逐漸減弱。若將標(biāo)本放在聚乙烯塑料袋中4℃保存，可延緩熒光減弱時(shí)間，防止封裱劑蒸發(fā)。</p><p> –用戊二醛或碳化二亞胺作為交聯(lián)劑通過(guò)功能基團(tuán)-NH2、-COOH等將半抗原結(jié)合到載體上。結(jié)合比例為5kDa結(jié)合5～25個(gè)分子的小肽。免疫細(xì)胞化學(xué)技術(shù)抗體(antibody,Ab)1、抗體的概念：免疫球蛋白·機(jī)體受到抗原刺激后，由漿細(xì)胞合成并分泌出一類具有與抗原發(fā)生特異性結(jié)合的球蛋白，被稱為抗體。–抗體主要存在于血清內(nèi)；–抗體都是免疫球蛋白，但免疫球蛋白并不一定都是抗體。–免疫球蛋白根據(jù)重鏈的結(jié)構(gòu)及抗原特異性不同分為五種，既IgG、IgD、IgE、IgA、IgM。2、免疫組化實(shí)驗(yàn)中常用的抗體：?jiǎn)慰寺】贵w和多克隆抗體。·單克隆抗體：是一個(gè)B淋巴細(xì)胞克隆分泌的抗體，是應(yīng)用細(xì)胞融合雜交瘤技術(shù)免疫動(dòng)物制備的。–特異性強(qiáng)、抗體產(chǎn)量高。·多克隆抗體：是將純化后的抗原直接免疫動(dòng)物后，從動(dòng)物血中所獲得的免疫血清，是多個(gè)B淋巴細(xì)胞克隆所產(chǎn)生的抗體混合物。–特異性低，會(huì)產(chǎn)生抗體的交叉反應(yīng)。–多克隆抗體廣泛應(yīng)用于石蠟包埋組織切片，可減少假陰性染色機(jī)會(huì)。–抗原與抗體的結(jié)合，要求量保持一定比例，當(dāng)抗原或抗體過(guò)量時(shí)，是不能聚合成大顆粒。3、抗體的制備——多克隆抗體的制備·動(dòng)物的選擇·佐劑(adjuvant)·免疫方法·免疫劑量·抗體效價(jià)的測(cè)定·放血或定期抽血。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="上海多靶點(diǎn)免疫組化染色服務(wù),免疫組化" src="http://tyunfile.71360.com/UploadFile/shchaorui/637124899748701172/8129573.png"></div><p> –缺點(diǎn)：對(duì)封閉牢固的抗原決定簇暴露不理想。(3)微波照射法·將玻片放入裝有抗原修復(fù)液的容器中，置微波爐加熱至95℃以上，持續(xù)l0～15分鐘，冷卻后，按免疫組化染色步驟進(jìn)行?！ご朔椒ㄓ捎谖⒉▓?chǎng)內(nèi)極性分子、離子高速運(yùn)動(dòng)，撞擊交聯(lián)的網(wǎng)鏈，使抗原異常的構(gòu)想恢復(fù)正常，且因分子運(yùn)動(dòng)產(chǎn)熱、效率高、時(shí)間短，對(duì)抗原再現(xiàn)效果好。(4)高壓加熱法暴露抗原·將玻片浸入抗原修復(fù)液內(nèi)，置高壓鍋中高壓2～3min，可取得極好的效果?！び捎诟邏合率軣峋鶆?，特別使用于大批量標(biāo)本的染色。(1)酸水解法·酸水解可使交聯(lián)斷裂、暴露抗原?！⒉Ｆ?mol/LHCl溶液中，室溫作用20min(溫度升高，作用時(shí)間縮短)。·此法能增強(qiáng)特異性染色，降低背景，但需注意水解過(guò)度將破壞抗原性及形態(tài)結(jié)構(gòu)。加熱法的注意事項(xiàng)·達(dá)到規(guī)定的溫度(92～95?C以上)；·維持一定的時(shí)間；·避免切片干涸(抗原可能完全丟失)；·加熱后必須經(jīng)過(guò)室溫自然冷卻20～30分鐘，使未折疊的蛋白分子鏈恢復(fù)天然構(gòu)型；·修復(fù)液：–**常用的是；–***研究表明堿性修復(fù)液更有效，推薦使用1mM的EDTA緩沖液()。4、載玻片的處理·抗原修復(fù)過(guò)程中，由于高溫、高壓等諸多固素的影響，極易造成脫片。為防預(yù)防脫發(fā)片，常用粘附劑處理載玻片。北京七色免疫組化檢測(cè)服務(wù)</p><p style="text-indent:25px">上海多靶點(diǎn)免疫組化染色服務(wù)</p><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)。上海多靶點(diǎn)免疫組化染色服務(wù)</p><p 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