鼠抗人CD7單克隆抗體

鼠抗人CD7單克隆抗體

廣州健侖生物科技有限公司

CD7是分子量為40kDa的細(xì)胞膜蛋白，為表達(dá)于胸腺細(xì)胞膜表面zui早和持續(xù)存在的T 細(xì)胞特異性抗原，主要存在于絕大多數(shù)T 細(xì)胞、胸腺細(xì)胞和大多數(shù)的NK細(xì)胞。此抗體可用于研究人的正常T 細(xì)胞，也可作為淋巴瘤及白血病的分類參考依據(jù)。

我司還提供其它進(jìn)口或國(guó)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(tuán)菌、化妝品檢測(cè)、食品安全檢測(cè)等試劑盒以及日本生研細(xì)菌分型診斷血清、德國(guó)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(hào)：MRQ-56

同型：IgG2b

適用組織：石蠟/冰凍

陽性對(duì)照：扁桃體

抗原修復(fù)：熱修復(fù)（EDTA）

抗體孵育時(shí)間：30-60min

一、基本原理
本方法先借助長(zhǎng)期混合淋巴細(xì)胞培養(yǎng)法獲得抗原特異性CTL，然后再進(jìn)行細(xì)胞毒試驗(yàn)。其原理為：外周血淋巴細(xì)胞包含針對(duì)不同抗原的特異性CTL克隆，在體外經(jīng)某一特定（或同種異體細(xì)胞）抗原刺激后，能識(shí)別該抗原的T細(xì)胞克隆被選擇性激活、增殖，而其他T細(xì)胞克隆則逐漸死亡；經(jīng)3~4次刺激后，存活的均為識(shí)別特異性MHC/抗原肽復(fù)合物的細(xì)胞，即抗原特異性CTL 。
二、試劑及材料
1. 抗原抗體C（Sigma）：用培養(yǎng)液或PBS配制300μg/ml。
2. 含20%的新生牛血清的RPMI 1640
3. EB病毒轉(zhuǎn)化的B淋巴母細(xì)胞株
三、操作方法
1. 特異性CTL的誘導(dǎo)和制備
①  取作者外周血分離PBMC，無血清1640洗兩遍，用含20%的新生牛血清的RPMI 1640調(diào)成1.5×106 /ml，置于24孔板中，于5% CO2 培養(yǎng)箱中4小時(shí)使單核細(xì)胞貼壁以去除之，然后收集細(xì)胞，計(jì)數(shù)；
②  取EB病毒轉(zhuǎn)化的B淋巴母細(xì)胞，加入抗原抗體C，zui終濃度為30μg/ml，于37℃水浴中作用30min，1000r/min離心10min，棄上清，沉淀細(xì)胞用1640液洗滌3次并計(jì)數(shù)；
③  取2×106 個(gè)PBL于24孔板中，加入5×104 （2.5%）個(gè)經(jīng)抗原抗體C處理（30mg/ml、30min）的自身、同種異體（其HLA-I類型別*不同）的EBV-LCL細(xì)胞作為刺激細(xì)胞，混勻，用*培養(yǎng)基（RPMI 1640）補(bǔ)總體積至2ml；
④  靜置于培養(yǎng)箱中；4d后半量換液，繼續(xù)培養(yǎng)3d；
⑤  離心收集細(xì)胞，取1×106 個(gè)反應(yīng)細(xì)胞，加入2×105個(gè)（20%）的刺激細(xì)胞，第三天加入重組IL-2，使終濃度為30U/ml；每三天半量換液一次并維持相同IL-2濃度。
⑥  每周按相同程序刺激效應(yīng)細(xì)胞一次，3~4次后，效應(yīng)細(xì)胞即為特異性CTL，可用于殺傷實(shí)驗(yàn)。

我司還提供其它進(jìn)口或國(guó)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(tuán)菌、化妝品檢測(cè)、食品安全檢測(cè)等試劑盒以及日本生研細(xì)菌分型診斷血清、德國(guó)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(chǎng)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(hào)二期2幢101-103室

First, the basic principle
In the method, antigen-specific CTL is obtained by long-term mixed lymphocyte culture method, and then cytotoxicity test is performed. The rationale is that peripheral blood lymphocytes contain specific CTL colonies against different antigens, which are selectively activated, proliferated, stimulated by a specific (or allogeneic) antigen in vitro, While other T cell clones gradually died. After 3 or 4 stimuli, the surviving cells that recognized the specific MHC / antigen peptide complex were antigen-specific CTLs.
Second, reagents and materials
1. Antibody Antibody C (Sigma): Prepare 300 μg / ml with culture medium or PBS.
2. RPMI 1640 with 20% newborn calf serum
EB virus transformed B lymphoblastoid cell line
Third, the operation method
1. Specific CTL induction and preparation
① PBMCs were isolated from the peripheral blood of the authors, washed twice with serum-free 1640, and transferred to 1.5 × 10 6 cells / ml with RPMI 1640 containing 20% ??fetal bovine serum in a 24-well plate in a 5% CO 2 incubator for 4 hours Adherent monocytes to remove, and then collect cells, counting;
② Take EB virus transformed B lymphoblasts, add antigen antibody C, the final concentration of 30μg / ml, 37 ℃ water bath for 30min, 1000r / min centrifugation 10min, the supernatant was discarded, the cells were washed with 1640 solution three times and count;
③ Take 2 × 106 PBL in 24-well plate, add 5 × 104 (2.5%) of the antigen and antibody C treatment (30mg / ml, 30min) of its own, allogeneic (HLA-I type compley different) Of EBV-LCL cells as stimulator cells, mix and make up to 2 ml with complete medium (RPMI 1640)
④ static placed in the incubator; 4d after half the amount of liquid, continue to c*te 3d;
⑤ Centrifugal collection of cells, take 1 × 106 reaction cells, add 2 × 105 (20%) of the stimulation of cells, the third day of recombinant IL-2, the final concentration of 30U / ml; And maintain the same IL-2 concentration.
⑥ weekly stimulation of effector cells by the same procedure once, 3 to 4 times, the effector cells are specific CTL, can be used for killing experiments.