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

URGENT! use of a conjugate as a primary antibody - (Feb/16/2010 )

Hi everyone!
I have a problem... I have no choice but to use an antibody conjugated with FITC as a primary antibody in a Western Blot, because it is not produced without label. Is it possible to detect this 1 congugate with a polyclonal serum labeled with Peroxidase? Have anyone tried? :(
comments and suggestions very wellcome!! :)

-clutz-

Seems like it would work, so long as your secondary HRP-labelled antibody is directed specifically against available epitopes on your primary antibody, and your primary antibody recognises epitopes on your antigen of interest...

-HomeBrew-

HomeBrew on Feb 16 2010, 01:57 PM said:

Seems like it would work, so long as your secondary HRP-labelled antibody is directed specifically against available epitopes on your primary antibody, and your primary antibody recognises epitopes on your antigen of interest...


I thought so. The FITC molecule is small and may block some epitopes on the 1 Ab, but leave some others exposed and the HRP-labelled antibody is directed against H+L chains so...Ill try.
Thanks HomeBrew!

-clutz-