We would like to determine by flow all the cell types in the thymus that present OVA peptide on MHC II in RIPmOVA mice to OT-II T cells. There is an antibody towards OVA:MHCI complexes, but cant find one for OVA:MHCII. Does anyone know of an antibody or TCR-multiplex that could detect this?
We would do a OT-II proliferation/CD69 expression assays, but would like to know which cells to sort and are also worried about low levels that would be undetectable with this assay.
Thanks!
Hi Todd,
I would suggest using a tetramer. It is even possible to get them for free from NIH already conjugated to fluorochromes (http://tetramer.yerkes.emory.edu/reagents/class-II-mhc?field_protein_value=&combine=OVA&field_sequence_value=). I would suggest using two tetramers for the same specificity but on different fluorochromes. This helps improve detection.
Thanks Zack. However, I was looking to see which APCs are presenting the peptide in rip-OVA mice, rather than Ag specific T-Cells. Any thoughts?
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