[Histonet] Immunocytochemistry question

[John C. Dennis]

Dear J Saltarrelli

Go ahead and do the experiment.  Run your protocol without primary and see 
what sort of non-specific signal there is to be seen.

Or does that open the empiricism vs rationalism can of worms?

John Carroll Dennis
Anatomy, Physiology, and Pharmacology
109 Greene Hall
Auburn University, AL  36849


On Thu, 29 Nov 2007, J. Saltarrelli wrote:

> 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
>
>
>
>
>
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