TY - JOUR
T1 - Accurate measurement of oxidized glutathione content of human, rabbit, and rat red blood cells and tissues
AU - Srivastava, Satish K.
AU - Beutler, Ernest
N1 - Funding Information:
‘This work was made possible by a supporting fund established in the name of Barry T. Leithhead Research Fellowship, and was supported, in part, by PHS Grant HE 07449 from the National Heart Instiute. and PHS Grant HD 01974 from the National Institute of Child Health and Human Development. 70
PY - 1968
Y1 - 1968
N2 - 1. 1. A method for the accurate measurement of oxidized glutathione (GSSG) in the erythrocytes and body tissues has been described. The oxidation of reduced glutathione to GSSG during protein precipitation by trichloroacetic acid is prevented by prior alkylation of the sulfhydryl groups with N-ethylmaleimide (NEM). Trichloroacetic acid and excess NEM are extracted with ether. The GSSG in the extract is measured enzymically using glutathione oxidoreductase. 2. 2. Oxidized glutathione content (average ± standard deviation) of human, rabbit, and rat erythrocytes has been shown to be 3.6 ± 1.4, 5.6 ± 0.7 and 6.1 ± 2.6 mμmoles/ml RBC, respectively. 3. 3. The GSSG content of rat liver, kidney, heart, lens, testes, and skeletal muscles is below the level of detection, i.e., 0.03 μmole/gm tissue.
AB - 1. 1. A method for the accurate measurement of oxidized glutathione (GSSG) in the erythrocytes and body tissues has been described. The oxidation of reduced glutathione to GSSG during protein precipitation by trichloroacetic acid is prevented by prior alkylation of the sulfhydryl groups with N-ethylmaleimide (NEM). Trichloroacetic acid and excess NEM are extracted with ether. The GSSG in the extract is measured enzymically using glutathione oxidoreductase. 2. 2. Oxidized glutathione content (average ± standard deviation) of human, rabbit, and rat erythrocytes has been shown to be 3.6 ± 1.4, 5.6 ± 0.7 and 6.1 ± 2.6 mμmoles/ml RBC, respectively. 3. 3. The GSSG content of rat liver, kidney, heart, lens, testes, and skeletal muscles is below the level of detection, i.e., 0.03 μmole/gm tissue.
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U2 - 10.1016/0003-2697(68)90082-1
DO - 10.1016/0003-2697(68)90082-1
M3 - Article
C2 - 5750729
AN - SCOPUS:0014428920
SN - 0003-2697
VL - 25
SP - 70
EP - 76
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - C
ER -