鼠抗人CD44v6單克隆抗體

廣州健侖生物科技有限公司

CD44V6是CD44家族的異構(gòu)體之一，該抗原主要存在于上皮細(xì)胞內(nèi)。有研究表明CD44V6的高表達(dá)與多種惡性腫瘤的侵襲相關(guān)，而其低表達(dá)則與肺低分化腺癌及鱗癌、低分化膀胱癌及低分化前列腺癌相關(guān)。主要用于各種上皮源性腫瘤的研究，如乳腺癌、結(jié)腸癌及肺癌等。

我司還提供其它進(jìn)口或國產(chǎn)試劑盒：登革熱、瘧疾、流感、A鏈球菌、合胞病毒、腮病毒、乙腦、寨卡、黃熱病、基孔肯雅熱、克錐蟲病、違禁品濫用、肺炎球菌、軍團菌、化妝品檢測、食品安全檢測等試劑盒以及日本生研細(xì)菌分型診斷血清、德國SiFin診斷血清、丹麥SSI診斷血清等產(chǎn)品。

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【產(chǎn)品介紹】

細(xì)胞定位：細(xì)胞膜

克隆號：VFF-7

同型：IgG1/K

適用組織：石蠟/冰凍

陽性對照：膀胱癌/肺鱗癌

抗原修復(fù)：熱修復(fù)（EDTA）

抗體孵育時間：30-60min

二、操作步驟
1. 收集凋亡細(xì)胞：取1～2×106 個細(xì)胞，離心后，立即用70%酒精（-20℃）固定2h；
2. 洗滌：取出酒精固定的細(xì)胞，1000r/min離心5min ，去掉上清液，PBS洗二次；
3. 裂解細(xì)胞：加400μl細(xì)胞裂解液，充分混勻再加蛋白酶K100μl，置65℃水浴消化2小時或過夜；
4. 蛋白處理：加75μl 8mol/L的醋酸鉀，4℃15min，再加750μl抗原抗體，充分混勻后，10000r/min離心10分鐘后，將上清移至一新的Eppendorf管中；
5. 沉淀DNA：加入750μl無水乙醇，上下輕柔顛倒混合，即可見乳白色沉淀，若不明顯時可置-20℃過夜，12000r/min離心10分鐘，去上清；
6. 洗滌DNA：加1ml70%乙醇，混勻，10000r/min 離心5min，去上清；
7. 溶解DNA：根據(jù) DAN 沉淀的大小，加一定量的蒸餾水或TE，37℃溶解；
8. 測定DNA濃度；
9. 2%瓊脂糖凝膠電泳80V 2小時；
三、結(jié)果判斷
出現(xiàn)梯狀電泳條帶，zui小的條帶為180～200 bp，其他的條帶為其整倍數(shù)大小。壞死細(xì)胞則出現(xiàn)彌散的電泳條帶，無清晰可見的條帶。正常細(xì)胞DNA基因條帶因分子量大，遷移距離短，故停留在加樣孔附近。
轉(zhuǎn)染 ，是將外源性基因?qū)爰?xì)胞內(nèi)的一種專門技術(shù)。隨著基因與蛋白功能研究的深入，轉(zhuǎn)染目前已成為實驗室工作中經(jīng)常涉及的基本方法。轉(zhuǎn)染大致可分為物理介導(dǎo)、化學(xué)介導(dǎo)和生物介導(dǎo)三類途徑。電穿孔法、顯微注射和基因槍屬于通過物理方法將基因?qū)爰?xì)胞的范例；化學(xué)介導(dǎo)方法很多，如經(jīng)典的磷酸鈣共沉淀法、脂質(zhì)體轉(zhuǎn)染方法、和多種陽離子物質(zhì)介導(dǎo)的技術(shù)；生物介導(dǎo)方法，有較為原始的原生質(zhì)體轉(zhuǎn)染，和現(xiàn)在比較多見的各種病毒介導(dǎo)的轉(zhuǎn)染技術(shù)。

我司還提供其它進(jìn)口或國產(chǎn)試劑盒：登革熱、瘧疾、流感、A鏈球菌、合胞病毒、腮病毒、乙腦、寨卡、黃熱病、基孔肯雅熱、克錐蟲病、違禁品濫用、肺炎球菌、軍團菌、化妝品檢測、食品安全檢測等試劑盒以及日本生研細(xì)菌分型診斷血清、德國SiFin診斷血清、丹麥SSI診斷血清等產(chǎn)品。

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【公司名稱】 廣州健侖生物科技有限公司
【市場部】     歐

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【騰訊  】 
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號二期2幢101-103室

Second, the steps
1. Collect apoptotic cells: Take 1 ~ 2 × 106 cells, centrifuge, immediay with 70% alcohol (-20 ℃) ??fixed 2h;
2. Washing: remove the fixed cells of alcohol, centrifuge at 1000r / min for 5min, remove the supernatant and wash twice with PBS;
3. Lysis of cells: add 400μl cell lysis solution, mix well with protease K100μl, set 65 ℃ water bath for 2 hours or overnight;
4. Protein processing: add 75μl of 8mol / L potassium acetate, 4 ℃ 15min, add 750μl of antigen and antibody, mix well and centrifuge at 10000r / min for 10 minutes, then transfer the supernatant to a new Eppendorf tube;
5. Precipitated DNA: add 750μl of absolute ethanol, gently mix up and down, milky white precipitate can be seen, if not obvious can be set at -20 ℃ overnight, 12000r / min for 10 minutes to the supernatant;
6. Wash DNA: Add 1ml70% ethanol, mix, 10000r / min centrifugation 5min, to the supernatant;
7. Dissolved DNA: According to the size of DAN precipitation, plus a certain amount of distilled water or TE, dissolved at 37 ℃;
8. Determination of DNA concentration;
9. 2% agarose gel electrophoresis 80V 2 hours;
Third, the result to judge
The ladder electrophoresis bands, the smallest band 180 ~ 200 bp, the other bands for the whole multiple size. Necrotic cells appear dispersed electrophoresis bands, no clear visible bands. Normal cell DNA gene band because of large molecular weight, migration distance is short, so stay in the sample hole near.
Transfection is a specialized technique for introducing exogenous genes into cells. With the further study of gene and protein function, transfection has become the basic method often involved in laboratory work. Transfection can be roughly divided into physical-mediated, chemical-mediated and bio-mediated three types of pathways. Electroporation, microinjection, and gene gun belong to the paradigm of gene transfer into cells by physical methods; there are many chemical mediation methods, such as classical calcium phosphate coprecipitation, liposome transfection methods, and various cationic species-mediated Technology; bio-mediated method, a more primitive protoplast transfection, and now more common various virus-mediated transfection technology.