Protocol Online logo
Top : New Forum Archives (2009-): : Immunology

No IgG Signals in immunostaining - urgent (Apr/10/2010 )

I am doing immunostaining with purified IgG from rabbit, can anyone give me explanation why there is no signal in my staining despite it shares reactivity with the protein that I stained, my ELISA shows positive titers but not very high.
another question if ELISA is positive does that mean the staining either will be positive?

-bluebird-

bluebird on Apr 10 2010, 10:56 PM said:

I am doing immunostaining with purified IgG from rabbit, can anyone give me explanation why there is no signal in my staining despite it shares reactivity with the protein that I stained, my ELISA shows positive titers but not very high.
another question if ELISA is positive does that mean the staining either will be positive?


Can be hundred of reasons.
First, which kind of staining are you doing exactly?
Second, have you inclued positive control?
and last, what does it mean "not very high signal in Elisa"?

-nicolas-

are you immunostaining a blot from sds-page? if so then your antibody is probably good for native protein but not denatured protein.

-mdfenko-