URGENT! use of a conjugate as a primary antibody - (Feb/16/2010 )
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 1º congugate with a polyclonal serum labeled with Peroxidase? Have anyone tried?
comments and suggestions very wellcome!!
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 on Feb 16 2010, 01:57 PM said:
I thought so. The FITC molecule is small and may block some epitopes on the 1º Ab, but leave some others exposed and the HRP-labelled antibody is directed against H+L chains so...I´ll try.