Glutathione, glutathione disulfide, and S-glutathionylated proteins in cell cultures
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- 作者：Daniela Giustarinia ; giustarini@unisi.it" class="auth_mail" title="E-mail the corresponding author ; Federico Galvagnib ; Anna Teseic ; Alberto Farolfid ; Michele Zanonic ; Sara Pignattac ; Aldo Milzanie ; Ilaria M. Maronef ; Isabella Dalle-Donnee ; Romina Nassinif ; Ranieri Rossia
- 关键词：actinSSG ; S-glutathionylated actin ; BAEC ; bovine aortic endothelial cells ; BCNU ; 1 ; 3-bis(2-chloroethyl)-1-nitrosourea ; DMEM ; Dulbecco&rsquo ; s modified eagle medium ; DTT ; dithiothreitol ; EMEM ; Eagle's minimum essential medium ; FBS ; fetal bovine serum ; GR ; glutathione disulfide reductase ; GSH ; glutathione ; GSSG ; glutathione disulfide ; HUVEC ; human umbilical vein endothelial cells ; mBrB ; monobromobimane ; NEM ; N-ethylmaleimide ; PBS ; phosphate buffered saline solution ; PSSG ; S-glutathionylated protei
- 刊名：Free Radical Biology and Medicine
- 出版年：2015
- 出版时间：December 2015
- 年：2015
- 卷：89
- 期：Complete
- 页码：972-981
- 全文大小：1183 K
文摘
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A new protocol for the reliable measurement of PSSG, GSSG and GSH in cell cultures is proposed.
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The protocol is based upon the inclusion of N-ethylmaleimide (NEM).
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The absence of NEM results in artificial increase of both GSSG and PSSG levels of one order of magnitude.
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GSSG is only increased at the highest concentration of disulfiram (200 μM) in most cell lines.
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PSSG increased at the lowest concentration of disulfiram (10 μM) in most cell lines.