One more thing. What do you use as a control? Isotype? No primary? Adam
On Sun, Jan 10, 2010 at 6:29 PM, Adam . <[email protected]> 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 <[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
