RE: normal human serum

From:Abizar Lakdawalla

RE: normal human serum

Hi Carol, there are two parts to this response the first is technical and the second is commercial.

About doing IHC with human antibody on human tissues,
the best approach is to use the same approach that many mouse-on-mouse kits use.
This is to prelabel the antibody with biotin-tagged proteins (usually an anti-antibody Fab fragment).
The procedure is:
To your prediluted antibody prep add an anti-human biotinylated Fab or related proteins (like anti-human Fc receptors). Incubate to allow binding to the primary antibody. Unrelated proteins (Albumin, etc.) will not be labeled. Excess labeled Fab fragments in the mixture are then adsorbed by adding human serum or purified human Fc fragments (preferred) to the antibody mixture.

The labeled antibody is then applied to the section. The primary antibody will bind to its antigen, but this primary antibody now has biotin stuck onto it. The bound antibody is detected by the addition of streptavidin-enzyme conjugate. No secondary antibody is required in this procedure. It is pretty difficult to get background staining by this procedure though the method may not work with all human or humanized antibodies.

Now the commercial stuff,
we have performed human on human ihc services with pretty good success and have created a human-on-human kit based on our experience with the human-on-human services as well as the mouse-on-mouse iso-IHC kit that we sell so if you are interested ...


> -----Original Message-----
> From: Carol Chang []
> Sent: Friday, January 25, 2002 10:59 AM
> To:
> Subject: normal human serum
> Thank you all for the response of anti-human secondary
> antibody request.
> But I got another problem I need your help. I am trying to do
> human Ab on
> human tissue IHC. So I also need normal human serum to block
> the secondary
> Ab before the complex is applied to sample. Could you tell me
> what's the
> appropriate and safe way to get normal human serum except
> getting it from
> our own body that I am scared to do?
> Thank you and the histonet is always the place I can rely on.
> Carol
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