use of a bacterial protease inhibitor cocktail with mammalia

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use of a bacterial protease inhibitor cocktail with mammalia

Post by rpwdoc » Tue Aug 26, 2014 10:08 pm

Dear fellow biochemists,

I need some advice on Western Blotting, more specifically the use of certain protease inhibitors with the RIPA cell lysis buffer and protease inhibitor cocktail. A Millipore technical representative recommended my using "539134 | Protease Inhibitor Cocktail Set III, EDTA-Free - Calbiochem" as a good general protease inhibitor cocktail.

I have another protease inhibitor cocktail which includes: AEBSF, bestatin, E-64, and pepstatin A (E.coli protease inhibitor cocktail) but it does not include the aprotinin or the leupeptin inhibitors. Is there a risk of proteolysis in a mammalian cell lysate without these latter 2 inhibitors? The specific cell types which we are using are: HUVEC (human umbilical vein endothelial cells) and HT-29 adenocarcinoma cell line.

Dr Robert Weinberg
Dept of Biology, MIT
email: [email protected]
fax: 617-977-0902

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Post by JackBean » Sat Nov 08, 2014 5:18 pm

well, it would be advisable to add them

Cis or trans? That's what matters.

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