164
edits
Changes
added different RIPA volume for WAT
#Weigh frozen tissue samples, only need 20-50 mg of tissue. If there is too much cut it off and return the extra tissue to the -80. Record the weight of each tissue.
#Cool the centrifuge to 4C.
#Add 20 uL/mg of RIPA (keep on ice) or other buffer to tissue (400-1000 uL)for liver and muscle. Add 2-3uL/mg of RIPA for WAT.
#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.