[Cell-biology] Immunofluorescence with two rabbit antibodies together.

[Morgan Xu]

   Dear all,
   
   Has anybody ever tried immunofluorescence with two primary antibodies
   both developed from rabbits?
   
   One of the antibodies is conjugated with Alexa 488 directly. I am
   planning to apply the non-conjugated primary and the corresponding
   secondary first. Then fix it with paraformaldehyde. And last applying
   the conjugated primary.
   
   The concern is how well the secondary antibody will be held by
   fixation. Also how well the Alexa dye will stand the paraformaldehyde
   fixation.
   
   Any suggestion will be appreciated. Thanks.
   
   Morgan