谷胱甘肽抗體Anti-Glutathione mAb操作使用文獻數據參考

產品詳情：

抗谷胱甘肽單克隆抗體:

目錄號101-A

產品描述:lgG2a小鼠谷胱甘肽-蛋白質復合物的單克隆抗體。特異性:谷胱甘肽

克隆:D8

來源:小鼠

純度:蛋自A純化的

緩沖液/組合物:1次聚苯硫醚pH7.2;0.01%鈉疊氮化物

濃度:1mg/ml; 可根據要求提供批量

存儲長期:-80°C

短期:4'C

應用:在非還原條件下檢測蛋白質-谷胱甘肽成癮者的蛋白質印跡。酶聯免疫吸附試驗(ELISA知識產權

稀釋:1:1000

運輸:濕冰

國內現貨授權備貨商:靶點科技

Virogen品牌谷胱甘肽抗體Anti-Glutathione助力蛋白二硫鍵異構酶（PDI）研究，美國Virogen貨號101-A見刊于JCI：

論文信息：

論文題目：

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

期刊名稱：J Clin Invest.

時間期卷：2008;118(3):1110-1122.

在線時間：2008年2月14日

DOI：doi.org/10.1172/JCI32376.

產品信息：

貨號：101-A

規格：100ug

品牌：Virogen

產地：美國

名稱：Anti-Glutathione mAb 100

授權現貨代理：Target Technology（靶點科技）

Virogen現貨谷胱甘肽抗體101-A助力蛋白質二硫鍵異構酶研究發表在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.

谷胱甘肽抗體Anti-Glutathione mAb操作使用文獻數據參考：

(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.