BCl-2 lysis buffer - BCL-2 lysis buffer recipe (Aug/27/2007 )

I am having a very difficult time extracting bcl-2 from my cells. I am using PC3 cells (a prostate cancer cell line) and other prostate cancer cell lines along with breast cancer cells. These cells are known to express this protein.
Does anyone have experience extracting this protein? and is there a recipe for a lysis buffer????

thanks so much!

Use ANY whole cell lysis buffer.
For example, 1% triton, 150 mM Nacl, 20 mM tris ph 7.5.


BCL-2 is small, therefore use a high % polyacrylamide gel eg. 12%.
Also, use alot of protein. 25 micrograms of whole cell lysate will give a very weak signal for BCL-2.
Try 100 micrograms.