[Histonet] Immunocytochemistry question

[J. Saltarrelli]

Hello all,

 

I have an ICC dilemma and was hoping that someone would be able to help.

 

I'm using ICC to test the phenotype of endothelial cells using anti-von
Willebrand factor.  The primary was produced in rabbit.  To try and
alleviate non-specific binding, as a control I used an isotype IgG that is
also produced in rabbit.  The secondary, FITC, is against rabbit.  Could
using an isotype IgG for blocking from the same species as the primary lead
to excessive non-specific binding??  Would running the test using IgG from
rat possibly reduce non-specific binding since the secondary is
anti-rabbit??

 

Thanks for all of your help

 

Thank you,

 

J. Saltarrelli, Ph.D.

Postdoctoral Research Fellow

Department of Bioengineering

Rice University