Tissue Protein Extraction Buffer (1X, 100 ml; Part No TEB-1)

Protocol (Keep solutions on ice):
Pulverize approximately 90 uL of tissue. Place tissue in a 1.5 ml round bottom microcentrifuge tube.
Add general phosphatase and protease inhibitor cocktails to 500 uL of ice-cold Tissue Protein Extraction Buffer.
Add 500 uL Tissue Protein Extraction buffer with inhibitors to pulverized tissue.
Homogenize tissue with a mini pestle-homogenizer using 15 strokes, 3 seconds/stroke on ice.
Centrifuge 12000g for 15 min at 4C.
Remove supernatant (without lipid layer) and transfer into another 1.5 ml tube.
Centrifuge again at 12000g for 15 min at 4C.
Transfer supernatant to another tube. The supernatant fraction contains the extracted proteins.
The Bradford assay can be used to quantitate extracted protein concentration.