washing after blocking PVDF filter - (Mar/27/2011 )

Im doing WB in this way, after the transfer of proteins, I block my filter in 5%milk diluted in PBStween 0.1%. then, I wash in PBStween and keep at 4°C. the day after, I do antibody incubation (actin) diluted in 5%milk -PBST. I got the signal of actin with some aspecifics.
Im wondering if washing the filter after blockin and washing step overnight is a problem!

Yes it could be a problem. You would be better off blocking it overnight.

bob1 on Mon Mar 28 00:01:59 2011 said:

ok, thanks,
in fact, im doing antibody incubation now I will see!