技术文章您现在的位置:首页 > 技术文章 > 谷胱甘肽抗体Anti-Glutathione mAb操作使用文献数据参考

谷胱甘肽抗体Anti-Glutathione mAb操作使用文献数据参考

更新时间:2025-01-19   点击次数:171次

产品详情:

抗谷胱甘肽单克隆抗体:

目录号101-A

产品描述:lgG2a小鼠谷胱甘肽-蛋白质复合物的单克隆抗体。特异性:谷胱甘肽

克隆:D8

来源:小鼠

纯度:蛋自A纯化的

缓冲液/组合物:1次聚苯硫醚pH7.2;0.01%钠叠氮化物

浓度:1mg/ml; 可根据要求提供批量

存储长期:-80°C

短期:4'C

应用:在非还原条件下检测蛋白质-谷胱甘肽成瘾者的蛋白质印迹。酶联免疫吸附试验(ELISA知识产权

稀释:1:1000

运输:湿冰

国内现货授权备货商:靶点科技


Virogen品牌谷胱甘肽抗体Anti-Glutathione助力蛋白二硫键异构酶(PDI)研究,美国Virogen货号101-A见刊于JCI:

论文信息:

论文题目:

Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation

期刊名称:J Clin Invest.

时间期卷:2008;118(3):1110-1122.

在线时间:2008年2月14日

DOI:doi.org/10.1172/JCI32376.

产品信息:

货号:101-A

规格:100ug

品牌:Virogen

产地:美国

名称:Anti-Glutathione mAb 100

授权现货代理:Target Technology(靶点科技)


Virogen现货谷胱甘肽抗体101-A助力蛋白质二硫键异构酶研究发表在JCI的材料和方法

Immunoprecipitation. For the detection of the constitutive glutathionylation of TF, solubilized monocytes were centrifuged 3 times at 3,000 g for 2 minutes at 4°C, and the supernatant was incubated with the monoclonal anti-GSH antibody (Virogen; or control IgG) overnight at 4°C. In other experiments, intact monocytes were incubated with anti-GSH antibody for 60 minutes at room temperature. Then, protein A Sepharose (Sigma-Aldrich; 50 μl) was added and the mixture incubated for 60 minutes at 4°C. Immunoprecipitated complexes were collected by centrifugation at 3,000 g for 2 minutes at 4°C. After washing 3 times with PBS, the pellets were incubated in SDS sample buffer (without reducing reagents) for 60 minutes at 20°C. After centrifugation, the supernatant was subjected to SDS-PAGE (7.5%), followed by electroblotting onto the nitrocellulose membrane. The membranes were then exposed to a monoclonal anti-TF antibody (or control IgG), followed by a horseradish peroxidase–conjugated anti-mouse IgG. The experiments were repeated 3 times.

TF glutathionylation. For the labeling of GSH with biotin, equimolar amounts (10 mM) of NHS-biotin (Pierce Biotechnology) and GSH dissolved in PBS were incubated at room temperature. The reaction was terminated after 1 hour, the residual biotin being quenched with ethanolamine (50 mM). To enable the protein incorporation of the labeled glutathione, the cells (or sTF) were incubated for 30–60 minutes with biotin-GSH (5 mM). Subsequently, the cells were washed and solubilized with 1% Triton X-100 in PBS. sTF was dialyzed using centricon filters. Then streptavidin beads (Sigma) were added to the biotinylated proteins of the cells (or sTF) and the mixture was incubated overnight. After sedimentation of the beads, they were washed twice with PBS containing 0.2% Triton X-100. The proteins were eluted in SDS sample buffer, separated by SDS-PAGE, transferred to nitrocellulose membranes, and the monoclonal anti-TF antibody was employed to detect the glutathionylated protein.

谷胱甘肽抗体Anti-Glutathione mAb操作使用文献数据参考:

image.png

(A) Basal glutathionylation of cell TF. Solubilized monocytes were immunoprecipitated with anti-GSH antibody or with isotype control antibody, as described in Methods. This was followed by western blotting with control antibody or anti-TF antibody.

(C) Glutathionylation attenuates TF activation. Cell lysis increases procoagulant activity, relative to intact cells, which is completely inhibited by anti-TF antibody. Increasing concentrations of DTT suppress TF activity. Glutathionylation in lysed cells was induced by preincubation with DTT (0.1 mM), followed by incubation with GSH (0.1 mM plus 0.1 mM diamide; 15 minutes). This increased the density of the band representing glutathionylated TF in immunoblots of lysed monocytes by 5.5-fold. Procoagulant activity of the lysates was determined by coagulation factor concentrate. *P < 0.05 versus control. n = 3–9.

靶点科技(北京)有限公司

靶点科技(北京)有限公司

地址:中关村生命科学园北清创意园2-4楼2层

© 2025 版权所有:靶点科技(北京)有限公司  备案号:京ICP备18027329号-2  总访问量:276776  站点地图  技术支持:化工仪器网  管理登陆