IHC with Nalm-6

Adam . via methods%40net.bio.net (by anonwums1 from gmail.com)
Sun Jan 10 19:29:53 EST 2010


Nalm6 cells are human pre-B cells so they should express the pre-B cell
receptor which has undergone VDJ recombination at the IgH locus but not the
IgL (and instead have a surrogate light chain). Naive B-cells should not
express IgG but rather IgM. Of course, IgM is really just a bunch of IgGs
stuck together, so there could be a chance that your anti-IgG secondary
reacts with that, but I think that's unlikely. Nalm-6 cells are human cells.
Has your secondary antibody been cross-adsorbed to human?

It's also possible that your problem is Fc mediated, as Nalm6 cells express
the Fc receptor (
http://www.nature.com/leu/journal/v16/n7/full/2402523a.html#fig2). You might
want to try an Fc block.

Adam

On Sun, Jan 10, 2010 at 4:33 PM, Scott Brown <SBrown from ccia.unsw.edu.au>wrote:

> Hi all,
>
> I have been trying to do some IHC and IF using Nalm-6 cells, on every
> occasion my controls look identical to the one I have used antibody on.
> Does anyone know if Nalm-6 cells express surface immunoglobins that my
> secondary may be binding to?
>
> Regards
>
> Scott
>
> Scott Brown
> PhD Candidate
> Molecular Carcinogenesis Program
> Children's Cancer Institute of Australia for Medical Research
> Lowy Cancer Research Centre, UNSW
> High Street (PO Box 81)
> RANDWICK NSW 2031
> AUSTRALIA
>
> Phone: +61 2 9382 1829
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