谷胱甘肽（glutathione，r-glutamyl cysteingl +glycine，GSH）是一種含γ-酰胺鍵和巰基的三肽，由谷氨酸、半胱氨酸及甘氨酸組成，存在于幾乎身體的每一個細胞。胱甘肽有還原型（GSH）和氧化型（GSSG）兩種形式，在生理條件下以還原型谷胱甘肽占絕大多數(shù)。谷胱甘肽還原酶可以催化兩型間的互變。Virogen的谷胱甘肽抗體，貨號101-A，可以檢測谷胱甘肽化蛋白質(zhì)復(fù)合物，當WB檢測時，需要在非還原條件下進行。具體WB檢測的數(shù)據(jù)，可以參考這篇文獻。

Antibodies

Glutathionylation was detected with the anti-glutathione antibody MAB5310 (Millipore). The immunoprecipitation of the glutathionylated proteins was carried out with a monoclonal anti-glutathioine (GSH) antibody (#101-A; Virogen). The Na,K-ATPase α1 isoform was immunoprecipitated with SC-21712 antibody (Santa Cruz Biotech., Dallas, TX) and immunedetected with the sc-28800 antibody (H-300; Santa Cruz Biotech.). All α subunits (α(all)) were immunoprecipitated with sc-28800 and immunodetected with the α5 antibody (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City).The β1 isoform was immunodetected with a polyclonal antibody generously provided by Dr. P.A Pedersen, University of Copenhagen. The β2 isoform was detected with the polyclonal antibody 06-1711 (Millipore).

Quantification of glutathionylation

Western blotting of homogenized muscle material has shown that many proteins are susceptible to glutathionylation (Mollica et al. 2012). To study the glutathionylation of Na,K-ATPase subunits it is therefore necessary to use a purification step to isolate the subunits. The level of glutathionylation was studied with two independent techniques.

Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數(shù)據(jù)參考,WB數(shù)據(jù)圖片：

The level of α and β subunits glutathionylation, the effects of exercise and terbutaline. The glutathionylated proteins were isolated with immunoprecipitation using anti-glutathione antibodies and Na,K-ATPase subunits quantified with Western blots of the immunoprecipitate (Method 2). (A) H; examples of the β1 subunits detected in the Western blots of two different homogenates (10 μg protein in each lane). IP: β1 subunits detected with anti-β1 antibodies in immunoprecipitates (from100 μg protein) obtained from the same two homogenates using the anti-GSH antibody.

The relative level of glutathionylated Na,K-ATPase α subunits (Method 1). (A) H; Western blot of muscle homogenate (10 μg protein per lane) labeled with the anti-GSH antibody. IP left: Western blots of two different immunoprecipitates (from 200 μg proteins) using the anti-α(all) antibody (H-300) and labeled on the gel with the anti-α(all) antibody α5. IP right: Western blots of the same two immunoprecipitates labeled with the anti-GSH antibody.