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→Basic Protocol
==Basic Protocol==
# Stimulate cells if necessary(i.e. insulin treatment for 10 min)# Wash cells 2x1mL (1mL per well for 12 well plates, 2mL per well for 6 well plates) with ice cold PBS -/- and aspirate
# Add 200uL [[Buffer:RIPA]] buffer and scrape cells
# Pipet into cold eppendorf tubes
# rotate end over end for 30 minutes at 4C 4oC to lyse
# Centrifuge 10 min at 13,000 RPM to clarify
# Transfer 150uL of lysate to fresh tube and do Bradford assay before add 150uL 2XSDS
# Load gel
