the sensitive amount of protein for western blotting - (Nov/02/2009 )

sorry, I am a fresh man of biology, could anybody tell me what is the sensitive amount of protein for western blot?
I purified GST-fusion protein and checked by SDS-PAGE, bands were clear ,
but for western blot I think I have to dilute the GSTs , now I need to know the sensitive amount of it so that I can determine how many times the protein to be diluted.
thank you everybody, best regards to you!

dllinpat on Nov 2 2009, 03:55 PM said:

it depends on the ptotein as well the the antibody titer value.
As a thumb rule, the western is atleast 10 to 20 times more sensitive than the SDS-PAGE (i am referring to the method to be used as a purity test.) If it is just for identity, 1000 times lesser amount can be loaded onto the WB. But also depends on the loading amount in SDS-PAGE
Good luck!!

thank you friend, I'm gonna try it now.

dllinpat on Nov 2 2009, 02:25 AM said:

You can choose to dilute your sample or the ab. There are some tables that compare sensitivities look at pierce supersignal.