whole cell extracts - (Sep/28/2006 )
Could anyone tell me how to obtain whole cell extracts from frozen adipose tissue. I need to use these extracts for ELISAs.
I have done this with brain tissue and heart tissue, it should be similar. I just homogenized (either with a mortar and pestle or a teflon homogenzier or a polytron type homogenizer) in a standard tissue lysis buffer (maybe 50 mM HEPES or Tris with 150mM NaCl 5mM MgCl2... protease inhibitors etc... pH 7.5 or so) with a little detergent as well (depending on whether or not you want to include membrane proteins or lipid-associated proteins). Then spin down a sample use the sup in the ELISA.
Hope this helps,