I would like to check GSH, SOD activity from cell lysates. Can I use cell lysates stored at -20C prepared by RIPA buffer for western analysis?
Sure you can use that. What is the concentration of your cell lysates? Newez RIPA doesn't hinder any antioxidant enzymes mediated catalysis. Which method are you using to determine GSH and SOD?
I have not started GSH and SOD assays.but i had cell lysate prepared with RIPA.concentration around 1ug/ul.
Hi Srilatha, sure you can check GSH and SOD activity in your cell lysate, RIPA buffer does not interfere in enzyme assay. For these enzymes you do also in gel assay where you only need is the SDS eliminated in 12% PAGE (without SDS), which is good for in gel SOD/GSH activity assay and also use this lysate for SOD/GSH proteins level by western blotting.
Best.
Yes. You can definitely use cell lysate in RIPA buffer for Western blot, and GSH and SOD assay. In my experience just do not heat the sample processing for SOD or GSH assay, but for Western boiling of sample is required every time you run gel for better result.
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