ELISA antibody-use for WB? - (Jun/06/2007 )

I want to test by WB if a particular protein is upregulated after a drug treatment. The only antibody for this protein which I have found has been optimised for ELISA but not WB... This may be a very general question, but should it work for WB or what are the reasons why it wouldn't??

Thanks!
Aisling

It should work, just probably you would need more Ab's for WB.

it may work.

for a western blot the antibody needs to recognize an epitope that is exposed when the protein is denatured.

for an elisa the antibody needs to recognize an epitope that is exposed in the native protein.

a polyclonal will probably work.

a monoclonal may work.

you will need to determine for yourself, but keep in mind that the company has already told you what they know the antibody will work with.

That's great, thanks for your help.

Aisling

hi,
as medfenko said have a look in to the datasheet, if u donot find enough information then denature (Bioling) ur protein prior to the ELISA and see whether it works, if it works then use bit higher concentration of Ab for the westernblotting.

gud luk
sravan.

you will need to determine for yourself. Optimal antibody dilutions could be different between assays (ELISA or WB). The first thing to do is look for optimal dilution of your antibody in wb. For this, in our lab, we use an accutran-cross blot-system for cross blot from Schleicher&Schuell that is very useful.

hi,

Yep, it's like what you've been told.. Since it hasn't been tested, the only way to find out is to try it...

Do you have a sample of the protein you can use as a positive control?