[Histonet] dual fluorescent staining

[Ross Benik]

Hello everyone, 

 

I am working on a protocol for a dual fluorescent stain with two mouse
antibodies (one is IgG1 and the other is IgG2a).  We started by
performing the two stains sequentially with a serum free protein
blocking step in between, tagging one antibody with a 488 FITC labeled
secondary and a 568 Cy3 secondary on the other (both goat anti mouse of
course).  With this method we are getting a lot of background staining
so I was wondering if anyone who has experience in this sort of work if
you have a recommendation of a proper blocking solution between the
sequential stains?  I've heard about using Rodent Block Rat from Biocare
Medical or an un-conjugated goat anti mouse IgG secondary immunoglobulin
but I would like to hear from you all as well! :-)    

 

Thanks, 

Ross