Bruce

I work with rat brain.  I've found that background is frequently a
function of the primary antibody and how much "non-specific" affinity any
particular antibody has for rat brain.  I've also found that some cell
types will show signal if the tissue is not quenched.  Whether or not the
signal/lack of signal is caused by endogenous peroxidase-like activity or
if the peroxide/methanol treatment degrades the antigenicity of the
epitope, I haven't investigated.  I did try H2O2 in buffer a couple of
months ago and got a very different staining pattern than H2O2 in MetOH
with the same antibody and in fact the H2O2/buffer showed staining in a
cell population on the negative control (no primary) tissues.

The upshot is that you need to keep and eye on your controls and consider
the
behavior of the tissue with a particular antibody.

Beam me up!

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


On Mon, 5 Apr 1999, Bruce A Rasmussen wrote:

> 
> Is background staining caused by endogenous peroxidase mainly a blood
> confound? In other words with well perfused brain tissue is H202
> unnecessary or not worth the tissue damage and loss of immunoreactivity?
> Many thanks,
> Bruce
> 
> 
> 
> --------------------------------------
> Bruce Rasmussen
> Predoc Fellow in Experimental Neuropsychology
> The Krasnow Institute for Advanced Study
> George Mason University
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