RIPA Buffer

Protocol tips

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	- Total cell extracts were prepared by treating cells with RIPA buffer (50 mM Tris base, 150 mM NaCl, 2 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 10 mM NaF, 1% IGEPAL, 0.1% sodium sodecyl sulfate (SDS), 0.5% sodium deoxycholate) containing 0.5% protease and phosphatase inhibitor cocktail.

Protocol tips
- Total cell extracts were prepared by treating cells with RIPA buffer (50 mM Tris base, 150 mM NaCl, 2 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 10 mM NaF, 1% IGEPAL, 0.1% sodium sodecyl sulfate (SDS), 0.5% sodium deoxycholate) containing 0.5% protease and phosphatase inhibitor cocktail.

Publication protocol

Total cell extracts were prepared by treating cells with RIPA buffer (50 mM Tris base, 150 mM NaCl, 2 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 10 mM NaF, 1% IGEPAL, 0.1% sodium sodecyl sulfate (SDS), 0.5% sodium deoxycholate) containing 0.5% protease and phosphatase inhibitor cocktail. Lysates were clarified by centrifugation and the protein concentration of the supernatant was determined using the Bradford protein assay reagent. Twenty micrograms of total cell lysates were denatured in 1 × SDS loading dye containing dithiothreitol. For western blot analysis, equal amounts of proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and analyzed. Proteins on the gel were transferred onto a PVDF membrane.

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