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
Any suggestion will be appreciated. Thanks.