Kappa/lambda staining

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From:Dave Tacha <dtacha@ncal.verio.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
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> Kappa/lambda technique

There is quite an art in getting good kappa/lambda staining, especially
if tissues have been decalcified. Here are some tipsI've used and
learned over the past 10 years.

I use an Ionic Exchange Decal Unit for bone marrow biopsies.  This works
very fast, but is much more gentle than HCL or RDO.  After,
decalcification, I do not use antigen retrieval.  I not sure about
DECLERE.  It may not work well with certain types  of antigens or
antibodies.  I've heard a lot of mixed reports over the Histonet.  I
prefer trypsin or pepsin digestion.  I also block with 10% goat serum.
This seems to slightly improve staining.  I prefer to use monoclonals
versus polyclonals.  Sometimes, I  dilute my antibodies in a mixture of
goat serum and casein. I  also develop the slides under the microscope
to try and determine end-point staining.  I prefer to use AEC as a
chromogen.  You can differentiate AEC (before hematoxylin staining) in
95% alcohol.  Then wash in water and observe the staining under the
microscope.  I repeat the steps, if necessary.  This technique really
helps improve contrast and reduces some of the background staining
caused by circulating antibodies.
I hope this has helped you.

If you want more information about the Ion Exchange Unit, e-mail me at

David Tacha HTL (ASCP)

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