In article <3cdoqr$r3k at threed.uchc.edu> wtrymbul at neuron.uchc.edu writes:
>In article <ferbas.5.00122E4A at cyto.medsch.ucla.edu>, ferbas at cyto.medsch.ucla.edu (John Ferbas) writes:
>>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.
>>===========================================================================================
>>John Ferbas Ph.D. Ferbas at cyto.medsch.ucla.edu>>UCLA MED/Cellular Immunology & Cytometry Voice: (310) 825 6340
>>12-236 Factor Bldg. FAX: (310) 794 2145
>>10833 Le Conte Ave.
>>Los Angeles CA 90024-1745
>>Dear John,
> I am a graduate student working on HLA class I structure. Is it
>possibe that W6/32 is not blocking your CTL epitope? Is your CTL response
>Allo or syngeneic? It is possible that an allo CTL is responding to
>another class I epitope unrelated to the W6/32 epitope and may not be
>sterically hindered. Why not try some other monomorphic determinant
>monoclonals such as TP 25.99 or some of the B2m specific monoclonals such
>as BBM 1 or L368.
>hope this was helpful.
>CPT
Dear John,
I am also going to be working with W6/32. I assume you're using it in
the effector phase of the CML. What concentrations are you using to
saturate the epitopes? What's your exact experimental technique? Did
you throw in nonspecific Ab? W6/32 should react w/ most HLA class
I's...Hmmm...
--
Derek Chan
dvchan at netcom.com
