RE: [Histonet] Kappa, Lambda

From:Rene J Buesa (by way of histonet)

What I will propose you probably will be an "uphill battle" with your
pathologist, but the use of bone marrow a problem because they have
been decalcified (presumably with a gentle EDTA protocol) and they are
small and one of the things I always found very frustrating was to have a
small control that will not last.
I always used tonsil as control and you will have to try to convince your
pathologist that if a tonsil is kappa+ and lambda+ the procedure is OK and
will detect K & L+ cells anywhere.
I always used monoclonal (mouse) Ig from DAKO at 1:25 dilution with HIER
with citrate pH6 in a steamer. The engogenous peroxidase was quenched with
3% H2O2 X 5 minutes after HIER.
Additional word of caution: controls initially + for K and L tend to weaken
when the slides are kept for 4-5 weeks. According with the control slides
you need, cut just enough slides for 1 week only.
Rene J.

"Sebree Linda A."  wrote:
Monoclonal K and L will almost always be cleaner than polyclonal. Also,
I would select several bm bxs for optimizing since some specimens are
just cleaner (or dirtier) than others. Hope this helps.

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
600 Highland Ave.
Madison, WI 53792
FAX: (608)262-7174

-----Original Message-----
[] On Behalf Of Jesus
Sent: Thursday, October 20, 2005 12:36 PM
Subject: [Histonet] Kappa, Lambda

I have a great question out there for the IHC people. Here in the lab
we are trying to work up our Kappa and lambda. The problem that most
people see is the excessive background staining that comes with these
two antibodies. we are treating our tissue in a H2O2 solution just
before we run them on the IHC stainer (Ventanna). Here is the kicker in
working them up we are using Bone marrow for control, because our
pathologists would like ,control that is similar to the disease process
they are looking for. Now one specimen control stains beautiful for
Kappa and the other specimen control stains beautiful for Lambda. This
brings me to the conclusion that the stain is working and also that
different specimens no matter what stain differently. Can someone give
me a universal way to make this easier on us. I know we need to block
the endogenous peroxidase, and we are using H2O2.. is there anything
that we can do better or is there another method for us to treat the

Jesus Ellin
Yuma Regional Medical Center

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