high antibody binding non specifically - (Jan/14/2006 )

hey

i'm trying to use an antibody to measure protein bindings of s. pyogenes pathogen, and i'm using bsa as my negative (which is what everyone uses)

the problem is even in the strains that i know don't bind these proteins the bsa result is the same as the result for my proteins (ie i'm getting a lot of non-specific binding)

does any one know of an alternative to bsa as a negative control for protein binding in assays?

thanks

-enterokinase-

QUOTE (enterokinase @ Jan 14 2006, 08:29 AM)

Other possibilities are casein or fat-free milk.
Perhaps your antibody is too concentrated. Increasing the volume of washing solutions might help, too.
Good luck.

-sergechampetier-

increase NaCl concentration in wash buffer (up to 500 mM)
addition of 0.05% tween-20 and 0.2% triton X-100 could improve wash

-laurence-

QUOTE (enterokinase @ Jan 14 2006, 11:29 AM)

You could try Prionex, a BSA alternative which is a highly purified collagen peptide fraction from pig skin. You can find it at

Centerchem website

-lightsup-