I have a frozen tube of protein extraction from 1 cm2 of bovine muscle,i want to add a protease inhibitor cocktail to this tube without thawing the protein.
I had stock solution from inhibitor cocktail(1 tablet dissolved in 10 ml PBS buffer),how much volume from this stock i must add to my frozen tube?
And how can i estimate the initial fluid volume in the tube without thawing?
I'm not sure how you can guess the volume without thawing other than using markings on the tube(if there are any) to estimate. But for the protease inhibitors, I have made 10X stocks by dissolving the tablet in 1 mL before instead of 10mL and then adding the 10X solution to your sample to make a total of 1X PI.
Yep you want to use it at 1X. I also have a protocol for a home-made 100X protease inhibitor cocktail I will share with you if you'd like, but you'd have to have all the stock PI's
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