鼠抗人CD7单克隆抗体

广州健仑生物科技有限公司

CD7是分子量为40kDa的细胞膜蛋白，为表达于胸腺细胞膜表面zui早和持续存在的T 细胞特异性抗原，主要存在于绝大多数T 细胞、胸腺细胞和大多数的NK细胞。此抗体可用于研究人的正常T 细胞，也可作为淋巴瘤及白血病的分类参考依据。

我司还提供其它进口或国产试剂盒：登革热、疟疾、流感、A链球菌、合胞病毒、腮病毒、乙脑、寨卡、黄热病、基孔肯雅热、克锥虫病、违禁品滥用、肺炎球菌、军团菌、化妆品检测、食品安全检测等试剂盒以及日本生研细菌分型诊断血清、德国SiFin诊断血清、丹麦SSI诊断血清等产品。

欢迎咨询

欢迎咨询

【产品介绍】

细胞定位：细胞膜

克隆号：MRQ-56

同型：IgG2b

适用组织：石蜡/冰冻

阳性对照：扁桃体

抗原修复：热修复（EDTA）

抗体孵育时间：30-60min

一、基本原理
本方法先借助长期混合淋巴细胞培养法获得抗原特异性CTL，然后再进行细胞毒试验。其原理为：外周血淋巴细胞包含针对不同抗原的特异性CTL克隆，在体外经某一特定（或同种异体细胞）抗原刺激后，能识别该抗原的T细胞克隆被选择性激活、增殖，而其他T细胞克隆则逐渐死亡；经3~4次刺激后，存活的均为识别特异性MHC/抗原肽复合物的细胞，即抗原特异性CTL 。
二、试剂及材料
1. 抗原抗体C（Sigma）：用培养液或PBS配制300μg/ml。
2. 含20%的新生牛血清的RPMI 1640
3. EB病毒转化的B淋巴母细胞株
三、操作方法
1. 特异性CTL的诱导和制备
①  取作者外周血分离PBMC，无血清1640洗两遍，用含20%的新生牛血清的RPMI 1640调成1.5×106 /ml，置于24孔板中，于5% CO2 培养箱中4小时使单核细胞贴壁以去除之，然后收集细胞，计数；
②  取EB病毒转化的B淋巴母细胞，加入抗原抗体C，zui终浓度为30μg/ml，于37℃水浴中作用30min，1000r/min离心10min，弃上清，沉淀细胞用1640液洗涤3次并计数；
③  取2×106 个PBL于24孔板中，加入5×104 （2.5%）个经抗原抗体C处理（30mg/ml、30min）的自身、同种异体（其HLA-I类型别*不同）的EBV-LCL细胞作为刺激细胞，混匀，用*培养基（RPMI 1640）补总体积至2ml；
④  静置于培养箱中；4d后半量换液，继续培养3d；
⑤  离心收集细胞，取1×106 个反应细胞，加入2×105个（20%）的刺激细胞，第三天加入重组IL-2，使终浓度为30U/ml；每三天半量换液一次并维持相同IL-2浓度。
⑥  每周按相同程序刺激效应细胞一次，3~4次后，效应细胞即为特异性CTL，可用于杀伤实验。

我司还提供其它进口或国产试剂盒：登革热、疟疾、流感、A链球菌、合胞病毒、腮病毒、乙脑、寨卡、黄热病、基孔肯雅热、克锥虫病、违禁品滥用、肺炎球菌、军团菌、化妆品检测、食品安全检测等试剂盒以及日本生研细菌分型诊断血清、德国SiFin诊断血清、丹麦SSI诊断血清等产品。

想了解更多的产品及服务请扫描下方二维码：

【公司名称】 广州健仑生物科技有限公司
【市场部】     欧

【】 
【腾讯  】 
【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期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.