Protein Extraction from Human Tissue/Tumor Samples - (Jun/03/2009 )
I am curious if anyone has suggestions for ways to extract protein from human frozen tumor samples. I was told just to use a cold mortar and pestle to powder the tissue, is this correct? Then I can proceed as usually with lysis buffer in order to obtain lysates for western blotting?
Also, if anyone has an estimate of how much tumor sample you need for western blotting it would be very helpful. My lab is having samples sent, and we need to know how much to request. For example, XXX mg?
Thanks so much!
As to the homogenization, you could do it with a pestle, but you're homogenization will likely not be very complete. Alternatively, and especially if tissue homogenization is something that you and / or your lab will be doing often, you should try a Bullet Blender. Your homogenizations will be bar more complete and consistent, and as a result you'll get better results and require less tissue.
As to how much tissue you need for a western... That's highly dependent on how abundant your protein is. What protein are you probing for? As a general rule, a few mg should be more than enough.
If there's anything else I can help you with, let me know.