Hello! I'm using your protocol but I am confused about the standard...

MB
Minh Hoang Bui
Jun 3, 2022
protocol Protocol: Fluorometric Estimation of Glutathione in Cultured Microglial Cell Lysate
Hello! I'm using your protocol but I am confused about the standard preparation. Cause by the way you mentioned in the protocol, the concentration of GSH (M = 303.77 g) should be 26 nmol/mol (not 26.4) and GSSG (M = 612.6) should be 13 nmol/ml (not 26.4 nmol/ml).
I also wonder how lon
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Vikas Singh Author Answered Jun 12, 2022

Immunotoxicology Laboratory, Food Drug and Chemical Toxicology Group and Nanotherapeutics & Nanomaterial Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), 31-Vishvigyan Bhawan, India

Hi Minh Hoang Bui
Please go through the fig 1 and fig.2 of the paper. This method detects GSH because OPT can only react with free sulfahydrl group of GSH. In case of GSSG detection, it is hydrolyzed to GSH at high pH12. Therefore GSSG standard solution has concentration of 26 nmole/ml rather than 13 nmole/ml. I hope this answers your question.
After mixing solution A and Solution B KPE buffer should be used within a week.

Thanks
Vikas
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