IHC with Nalm-6

Adam . via methods%40net.bio.net (by anonwums1 from gmail.com)
Sun Jan 10 19:37:32 EST 2010


One more thing. What do you use as a control? Isotype? No primary?

Adam

On Sun, Jan 10, 2010 at 6:29 PM, Adam . <anonwums1 from gmail.com> wrote:

> 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
>> Fax: +61 2 9382 1850
>> Email: sbrown from ccia.unsw.edu.au
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