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.

Zenobia
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
>suggestions???????
>
>Sherry Smith
>Presbyterian Hospital
>Charlotte, NC
>
>
>This message and any included attachments are from NOVANT HEALTH INC.
>and are intended only for the addressee(s). The information contained
>herein may include trade secrets or privileged or otherwise confidential
>information. Unauthorized review, forwarding, printing, copying,
>distributing, or using such information is strictly prohibited
>and may be unlawful. If you received this message in error, or have
>reason to believe you are not authorized to receive it, please promptly
>delete this message and notify the sender by e-mail.
>
>Thank you.
>
>

Zenobia Haffajee
Anatomical Pathology 
HAPS John Hunter Hospital
dpeppera@mail.newcastle.edu.au
Tel :(02) 4921 4449
Fax :(02) 4921 4507




<< Previous Message | Next Message >>