Flow cytometry using anti-HLA class II antibody F3.3 Human leukocytes were stained with an isotype control (panel A) or the rabbit-chimeric version of F3.3 panel B) at a concentration of 1ug/ml for 30mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody and cells analysed on a FACSCanto flow cytometry.

Western Blot using anti-HLA class II histocompatibility antigen DR antibody F3.3 Human spleen (A), human tonsil (B) and Daudi cell lysate (C) samples (35ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of F3.3 at 0.01ug/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for HLA II DR is ~30kD, depending upon the exact DR isoform expressed (c.f. Klitz et al, PMID: 12974796). Successfully detected human HLA class II histocompatibility antigen DR in human spleen and tonsil lysate as well as Daudi cell lysate.