HLA class II monoclonal antibody, clone F3.3

Recommend Usage:
Flow Cytometry
Western Blot
The optimal working dilution should be determined by the end user.

Storage Instruction:
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.

Western Blot (Tissue lysate)
Western blot analysis of HLA class II monoclonal antibody, clone F3.3 (Cat # RAB01090).
Human spleen (A), human tonsil (B) and Daudi cell lysate (C) samples (35 ug protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit antibody (Cat # RAB01090) at 0.01 ug/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 (c.f. Klitz et al, PMID: 12974796). Cat # RAB01090 successfully detected human HLA class II histocompatibility antigen DR in human spleen and tonsil lysate as well as Daudi cell lysate.

Flow Cytometry
Flow-cytometry using HLA class II monoclonal antibody, clone F3.3 (Cat # RAB01090).
Daudi cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric antibody (Cat # RAB01090, blue line) at a concentration of 10 ug/mL for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody at 2 ug/mL and cells analyzed on a FACSCanto flow-cytometer.

Flow Cytometry
Flow-cytometry using HLA class II monoclonal antibody, clone F3.3 (Cat # RAB01090).
Human leukocytes were stained with an isotype control (panel A) or the rabbit-chimeric version of F3.3 (Cat # RAB01090, panel B) at a concentration of 1 ug/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.