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Preparing Cell Lysates

519 bytes added, 15:40, 11 May 2009
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type something hereMaterialsRIPA Buffer (for 10mL lysis buffer)Tris pH7.4 50mM 500uLNa Deoxycholate 0.25% 250uLNP-40 1% 1mLNaCl 150mM 371uLEDTA 1mM 20uLNaVO3 1mM 100uLNaF 1mM 20uLProtease Inhibitors 1 tab Basic Protocol# Stimulate cells if necessary# Wash cells 2x1mL with ice cold PBS -/- and aspirate# Add 200uL RIPA buffer and scrape cells# Pipet into cold eppendorf tubes# rotate end over end for 30 minutes at 4C to lyse# Centrifuge 10 min at 13,000 RPM to clarify# Transfer 150uL of lysate to fresh tube and add 150uL 2XSDS# Load gel
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