Re: Kappa and Lambda

From:Debbie Pepperal

Hi Sherry

there is quite a problem when it comes to Immunoglobulin staining for kappa
and lambda light chains. We have had some good staining; we use the Dako
antibodies  , both polyclonal. We use enzyme digestion plus MW antigen
retrieval in 4M Urea. You however will get some background staining with
regard to the degree fixation of the tissue. We do our staining manually
and use a PBS buffer with Tween 20 in it.

hope this helps a bit.

At 01:19 PM 04/10/2001 -0400, you wrote:
>Help!!  I am having a problem with background staining with Kappa and
>Lambda.  I know what the problem is but do not know how to solve it.  I am
>told that it is a IgG problem but how do you block out the background??  Our
>current method is as follows:  We use Cell Marque (monoclone) antibody.  No
>heat retrieval.  We use Protease II for 8min. and incubate for 32min.  It
>looks great except for the background staining.  Does anyone have any
>Sherry Smith
>Presbyterian Hospital
>Charlotte, NC
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Zenobia Haffajee
Anatomical Pathology 
HAPS John Hunter Hospital
Tel :(02) 4921 4449
Fax :(02) 4921 4507

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