Changes

Jump to: navigation, search

Preparation of Protein Lysates from Mouse Tissues

13 bytes added, 21:27, 26 July 2013
m
Protocol
#Centrifuge at 14 000 RPM at 4C for 10 min
#Remove supernatant to clean tube. If lysing fat, try to avoid the floating fat cake. If necessary respin to clarify
#Do a bradford assay using 1 uL of clarified lysate and label tube with correct concentration Follow Protein Lysate instructions for Bradford Assay (see [[Bradford Assay]])
#Prepare samples for gels by adding 800 ug protein to a final volume of 200 uL of lysis buffer. Add 200 uL of 2X loading buffer with B-ME to each lyaste. This will generate a 2 mg/mL protein solution in SDS Loading Buffer
#Heat samples with loading buffer at 95C for 5 mins
#Snap freeze remaining clarified lysate and SDS-PAGE lysates and store at -80
34
edits

Navigation menu