I will make extraction of muscle proteins for electrophoresis study. Does anyone have a suggestion for the concentration of protease inhibitors? Based on some studies, I'm thinking to use: benzamidine hydrochloride hydrate (1 mM), phenylmethanesulfonyl fluoride (PMSF) (1 mM) and EDTA (10 mM). Thank you very much
Hi Rafael, I would recommend using a commercial proteinase inhibitor cocktail that will inhibit 3 classes of common proteinases.
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Datasheet/5/p8340dat.pdf
If you add EDTA to the cocktail, then it will inhibit most metallo-proteinases as well.
It is better if you could incorporate Leupeptin , Pepstatin , Benzamidine , Antipain ,Chymostatin and also EDTA since muscles are calcium rich tissues.PMSF however is more useful if you need to block phosphatase activity.
above mentioned formulations are time consuming to make so Its better if you use Thermo Protease inhibitor cocktail Tablet or liquid.
Moreover the amount of protease inhibitors needed depends upon the amount of proteases in a given tissue and not as such the proteins.
Hope it helps.
Regards
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