Buffer A contained 50 mM tris-HCl (pH 7.5) and 0.1 mM EGTA. Lysis buffer was 50 mM tris-HCl (pH 7.5), 1 mM EGTA, 1 mM EDTA, 50 mM sodium fluoride, 5 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1% (w/v) Triton X-100, 0.27 M sucrose, 0.1% (v/v) 2-mercaptoethanol, and protease inhibitors (complete protease inhibitor cocktail tablets, Roche, 1 tablet per 50 ml). TBS-Tween buffer (TTBS) was tris-HCl (pH 7.5), 0.15 M NaCl, and 0.2% (v/v) Tween 20. SDS sample buffer was 1× NuPAGE LDS sample buffer (Invitrogen) containing 1% (v/v) 2-mercaptoethanol. Protein concentrations were determined following centrifugation of the lysate at 16,000g at 4°C for 20 min using the Bradford method with bovine serum albumin as the standard.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.