Reagents viz; Urea, thiourea, tris, and CHAPS used were of sisco laboratories make and DTT and Bradford reagent were Biorad make. Protein was quantified by quick start Bradford biorad make ragent method. Tissue lysis(1.5g) had been carried out in 10ml lysis buffer(# Urea 8M, thiourea 2M, tris 20mM, DTT 0.55M and CHAPS 4%). Different combinations were applied to maximize the protein release from tissue; pestle mortar for25 min., tissue homogenizer for 6min and ultrasonicator for 3 min. Lysate was centrifuged at 15000xg, at 4 degree C for thirty minutes and protein was estimated in supernatant by Bradford method. Table showed the protein yield (mg/ml). I also tried to check the conc. Of protein by diluting the lysate 5times in distilled water and in # urea buffer and compared with BSA standard in dw and # urea buffer