Substitute for BSA blocking in Westerns - options for blocking agents for phosphop (Aug/25/2003 )

What do you folks use for a blocking agent when running westerns for phosphorylated proteins? I am looking for an inexpensive alternative to BSA for MAPK expression analysis when using antibodies to phospho-Erk1 and Erk2.

try using 1% gelatin in TBST/PBST for blocking, it gives me equally good results. you can buy it from any grocery store..

4% powdered milk is used in the Upstate biotech anti P-Tyr protocol (4G10 antibody).

I've used 4% mlk for detection of P-CREB on PVDF.

i am using 1% skim milk TBS buffer

using 1% milk with no problem.