animal tissue extraction? - (Sep/01/2006 )

hi I want to check presence of specific protein in cancerous tissue of mice...
I dont know anything about how to process the tissue until getting a lysate to work with .. any body help me please

u want to make a protein lysate. I have used the follwing for neuronal tissues.

50 mM Tris 0.605 gms/ 100ml
150mM NaCl 0.87 gms/ 100ml
1% Triton X 100 1ml/ 100ml

+ protease inhibitor.

Add 100µl of buffer for a small piece 50-100mg of tissue in 4 ml of SNAP cap tube.(one should adjust vol. according to tissue size)

homogenise it

leave it on ice for 20min to lyse it

centrifue 15min at full speed

aliquot the supernatant and freeze in –80°C