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 <[email protected]>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: [email protected] > Web: www.ccia.org.au <http://www.ccia.org.au/> > Children's Cancer Institute Australia is the only independent medical > research institute in Australia solely devoted to research into the > causes, prevention and cure of childhood cancer. Our vision is to save > the lives of all children with cancer and eliminate their suffering. > The information contained in this message and any annexure is > confidential and intended only for the named recipient(s). If you have > received this message in error, please contact the sender immediately > and destroy the original message. > > > > Children's Cancer Institute Australia for Medical Research is the only > independent medical research institute in Australia devoted to research into > the causes, prevention, better treatment and ultimately a cure of childhood > cancer. Our vision is to save the lives of all children with cancer and to > eliminate their suffering. > > The information contained in this message and any annexure is confidential > and intended only for the named recipient(s). If you have received this > message in error please contact the sender immediately and destroy the > original message. > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
