[Histonet] Ki-67 (MIB1) Immunohistochemistry and neutrophillabeling

Wed Jun 2 12:00:34 CDT 2004

Thanks Patsy.  I agree with you.
Dave

David L. Krull
Scientist, Molecular & 
Ultrastructural Pathology
GSK Safety Assessment 9.2103
5 Moore Drive
RTP, NC 27709-3398

Phone: 919-315-2457
David.L.Krull <@t> GSK.com

"Patsy Ruegg" <pruegg <@t> colobio.com>

02-Jun-2004 12:26

        To:     david.l.krull <@t> gsk.com, ree3 <@t> leicester.ac.uk

        cc:     histonet <@t> lists.utsouthwestern.edu
        Subject:        RE: [Histonet] Ki-67 (MIB1) Immunohistochemistry and neutrophillabeling

I have always been taught to discount neutrophil staining in IHC, they 
seem
to stain with just about everything, not sure if this is unquenched
endogenous peroxidase or some other mechanism but we just don't count them
as positive results, even for the QIHC exam when we had to do an IHC stain
on tissue that was negative for a particular antibody to prove that it was
viable if we only got neutrophil staining and not vimentin or cytokeratin 
or
what ever, we did not count the neutrophil staining as evidence that the
tissue was still viable.
My two censt worth.
Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of
david.l.krull <@t> gsk.com
Sent: Wednesday, June 02, 2004 8:48 AM
To: ree3 <@t> leicester.ac.uk
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Ki-67 (MIB1) Immunohistochemistry and
neutrophillabeling

It's unlikely to be endogenous peroxidase.  I compared 3% hydrogen
peroxide(aq) - 10min with 3% hydrogen peroxide in methanol - 30min with
the same results; negative isotype control, positive neutrophils in the
DAB test section.  Note: Dako Biotin Blocking system was also used in the
protocol.

David L. Krull
Scientist, Molecular &
Ultrastructural Pathology
GSK Safety Assessment 9.2103
5 Moore Drive
RTP, NC 27709-3398

Phone: 919-315-2457
David.L.Krull <@t> GSK.com

"Edwards, R.E." <ree3 <@t> leicester.ac.uk>

02-Jun-2004 09:59

        To:     david.l.krull <@t> gsk.com

        cc:
        Subject:        RE: [Histonet] Ki-67 (MIB1) Immunohistochemistry 
and
neutrophil labeling

Endogenous peroxidase???????????????

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of
david.l.krull <@t> gsk.com
Sent: 02 June 2004 14:50
To: histonet <@t> lists.utsouthwestern.edu
Cc: charles.w.qualls <@t> gsk.com
Subject: [Histonet] Ki-67 (MIB1) Immunohistochemistry and neutrophil
labeling

We recently had a Cyno monkey study which required evaluation of cell
proliferation in formalin fixed, paraffin embedded liver sections.  During

the review of the slides we noticed that neutrophils were also positive in

addition to proliferating hepatocytes using DAB as the chromagen.  When I
switched to Vector VIP the neutrophils were negative.  Has anyone else
observed neutrophil labeling with Ki-67 using  DAB?  Could the DAB be
reacting with the neutrophils?

David L. Krull
Scientist, Molecular &
Ultrastructural Pathology
GSK Safety Assessment 9.2103
5 Moore Drive
RTP, NC 27709-3398

Phone: 919-315-2457
David.L.Krull <@t> GSK.com

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