Anti-HLA class II [F3.3]

Flow-cytometry using anti-HLA class II antibody F3.3 Human leukocytes were stained with an isotype control (orb256390, panel A) or the rabbit-chimeric version of F3.3 (orb411560, panel B) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using a AF488 conjugated donkey anti-rabbit antibody and cells analysed on a FACSCanto flow-cytometer.

Flow-cytometry using the anti-HLA II DR antibody F3.3 Daudi cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of F3.3 (orb411560, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.

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 (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of F3.3 (orb411560) at 0.01 µg/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 ~30 kDa, depending upon the exact DR isoform expressed. orb411560 successfully detected human HLA class II histocompatibility antigen DR in human spleen and tonsil lysate as well as Daudi cell lysate.