Sean Nair writes:
> Does anyone know of an RT-PCR kit for looking at the expression of T-cell
> Vbeta subsets.
>> Thanks for your help.
Sean:
To the best of my knowledge, there isn't one. However, the game is a
bit trickier than just designing primers and performing amplifications.
It is critical to have internal TCR standards to compensate for the
(dramatic) amplification efficiency differences between the various
V-beta amplicons. I wrote a book chapter on the subject a few years ago
(Spinella, DG and Robertson, JM, "Analysis of Human T-cell receptor
repertoires by PCR" in "The Polymerase Chain Reaction", KB Mullis, F
Ferre, and RA Gibbs, eds; 1994 Birkhauser, Boston). Read the paper and
if you find it convincing and want to do the analysis the same way, I
will send you the primers and internal reference standards (assuming
that it is human V-betas that you are working with -- I'm afraid I have
no mouse reagents). R&D Systems contacted me a while ago about the
possibility of making a kit from these reagents, but I guess they
decided that there wasn't enough market, so it never went anywhere.
Cheers.
--Dom Spinella
