I had assumed that John Ferbas was using HUMAN cells since he used W6/32
which we use as a strictly human antibody, but of course it cross reacts
with mouse cells (when they are grown in bovine serum). The method just
suggested is of course the classical approach, but using mabs is somewhat
easier than getting good cell lines at least sometimes.
Jeff Frelinger
ps see Klein book "Natural History of the MHC"
On Sat, 3 Dec 1994, antibody wrote:
> In article <Pine.3.89.9411301734.A300314-0100000 at UNCVX1.OIT.UNC.EDU>,
>JAF at UNCVX1.OIT.UNC.EDU wrote:
>> > try using mhc mismatached cells as well as a variety of other mabs.
> > W6/32 binds to the wrong side of class I.
> >
> >
> > On Tue, 29 Nov 1994, John Ferbas wrote:
> >
> > > I have used an anti-MHC class I antibody (W632) in an attempt to
> demonstrate
> > > MHC class I restriction in 2 CD8+ CTL clones, but have had no
> success. I know
> > > that the antibody is saturating the targets by FACS analysis. Also, the
> > > antibody is added before the assay and present throughout. Is there any
> > > alternative way to demonstrate class I restriction? Your comments are
> greatly
> > > appreciated.
> > >
>> Actually in my reply I totally forgot to mention that the most promising
> way to test for the restriction element is to use congenic MHC recombined
> mouse strains. Thats probably the classical, more work consuming way, but it
> gives you the right answer. Only if you have some clones coming from some
> bizarre H-2s than you might run into toubles to get the recombinants.
>> To look for mice strains with recombinant MHCs take a look at the article
> of Klein-J, Immunogenetics, I think Vol. 17, around page 700. "H-2 haplotypes,
> 2nd listing" or something like that.
>> Goog luck
>> --
> ******************************************************************
>> Joerg Kirberg EMAIL: joerg.kirberg at bii.ch> Basel Institute for Immunology FAX: 41-61-605 13 72
> Grenzacherstr. 487 PHONE: 41-61-605 12 77 (lab)
> Postfach
> CH-4005 Basel
>> Switzerland
>>
