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    • #17889
      rpwdoc
      Participant

      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

    • #115547
      JackBean
      Participant

      well, it would be advisable to add them

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