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Secondary AB blocking - (May/17/2012 )

Hi everyone,

I have a simple question, currently I'm having problem with my western blot with multiple bands. So now I trying to optimize it by:

- Overnight BSA blocking
- Decreased primary antibody concentration (5uL in 10mL) and increased in incubation time
- Decreased secondary antibody concentration (5uL in 10mL)

I am interested in blocking my secondary antibody (Goat vs Rabbit) with donkey serum (since its available in the lab). Is it possible since its a different animal?

Thanks for your time and attention..


yes, but you would be better off using serum from the animal in which the secondary antibody is raised (goat, in your case).

you can also prepare your primary antibody in the same medium.

we also add it to our block (bsa+goat serum).