RE: [Histonet] BKV background

From:"Favara, Cynthia \(NIH/NIAID\) [E]"

This is interesting to me as I was having a similar problem with an
antibody that I use frequently on murine brain and had cytoplasmic
staining in neurons but only on the outer edge of the tissue and the
outer portion of the brain [away from the midline]. I took different
blocks of brain tissue, same strain, same age but midline and did not
get the staining. I did not notice this staining of neurons when I
initially used this block. My simple thinking is that is may be what I
am calling an edge effect of fixation and or processing. Most outer
portion of tissue is exposed to fix and processing reagents for a bit
longer may result over processed tissue. In my experience over processed
tissue is more likely to have problems with artifacts. 

I am sure someone will have a different explanation!

Cynthia Favara
903 South 4th Street
Hamilton, MT 59840

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-----Original Message-----
From: Elaine Dooley [] 
Sent: Thursday, June 08, 2006 11:05 AM
Subject: [Histonet] BKV background

Dear Histonetters,

In the last couple of weeks we have been getting nuclear staining on
the edges of most of our kidney biopsies when we stain them for BKV. 
Our negative control is negative. (both the negative and the BKV slides
are treated with a protease digestion and are blocked for endogenous
biotin).  The control tissue we have is a large piece of autopsy kidney
positive for BKV.  It does not have nuclear staining on the edges.  In
the past we never seen nuclear staining on the edges of our needle
biopsies.  The processing area says they have not changed anything in
the processing of kidney needle biopsies.  We use Chemicons BKV large T
antigen antibody we have tried many different  dilutions but so far we
are still seeing the artifact.  Has anyone out there had this problem?

Elaine Dooley
Shands Teaching Hospital
325-265-0111 ext. 72117

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