[Histonet] Problems with secondary antibodies

From:Brian Dias

hi everyone,
    i'm trying to get a double label ICC to work using a Rbt polyclonal 
(against Glucocorticoid receptor) and a Ms monoclonal (against Tyrosine 
hydroxylase) primary combination. both of these work in single label runs 
using Biotinylated secondaries. however, when i switch to double 
fluorescent runs i run into the following problems:
1. my anti-Rbt FITC doesn't light up any cells at all (in single as well as 
double studies)(my anti-Ms Tx Red works just fine and i see some nice 
staining in both my single and double studies)
2. i thought this might be secondary problem, but when i use another 
Ployclonal Rbt (towards Tyrosine hydroxylase) and anti-Rbt FITC, i see some 
nice staining comparable to when i use the mouse monoclonal.
   any suggestions would be welcome.
thanks a ton


    Brian George Dias

    Graduate Student (Crews lab)            Office Phone #: 512-475-6738
    The University of Texas at 
Austin       E-mail:   brian.dias@mail.utexas.edu
    Institute for Neuroscience                  Website: 

    Patterson 56
    1 University Station Stop C0930
    Austin, TX 78712

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