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#Add 20 uL/mg of RIPA (keep on ice) or other buffer to tissue (400-1000 uL)
#Add a stainless steel bead and keep tissues on ice. Homogenize using Qiagen Tissue Lyser (3 min at 25Hz for WAT/Liver and 5 min at 50 Hz for Muscle).
#Remove stainless steel bead or transfer the homogenized tissue to a new tube. Keep tissue on ice.
#Centrifuge at 14 000 RPM at 4C for 10 min
#Remove supernatant and place into clean tube. If lysing fat, try to avoid the floating fat cake. If necessary respin to clarify
#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 85C for 5 2 mins
#Snap freeze remaining clarified lysate and SDS-PAGE lysates and store at -80