- Ab00185-1.1 Anti-HLA class II [F3.3]
- Mouse IgG1
- In Stock
- Ab00185-23.0 Anti-HLA class II [F3.3]
- Rabbit IgG
- In Stock
Recombinant monoclonal antibody to HLA class II. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma F3.3.
Alternative Name(s) of Target: HLA class II related epitope; HLA-DR
Specificity: Recognising anti-class II HLA molecules.
Application Notes: This antibody has been show to work by flow-cytometry and western blot. Dechant et al (PMID: 12393717) used a human IgA2-chimeric version of this antibody to study IgA-mediated cell killing.
Antibody first published in:
Elsässer D, Valerius T, Repp R, Weiner GJ, Deo Y, Kalden JR, van de Winkel JG, Stevenson GT, Glennie MJ, Gramatzki M. HLA class II as potential target antigen on malignant B cells for therapy with bispecific antibodies in combination with granulocyte colony-stimulating factor. Blood. 1996 May 1;87(9):3803-12. PMID:8611706
Note on publication: Describes the preparation and characterisation of anti human HLA class II antibody by of ADCC of G-CSF primed PMN expressing in the presence of F3.3 to HLA class II related epitope (HLA-DR).
Flow-cytometry using anti-HLA class II antibody F3.3 (Ab00185) Human leukocytes were stained with an isotype control (Ab00102-23.0, panel A) or the rabbit-chimeric version of F3.3 (Ab00185-23.0, 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 (ab150073) and cells analysed on a FACSCanto flow-cytometer.
Western Blot using anti-HLA class II histocompatibility antigen DR antibody F3.3 (Ab00185) Human spleen (A), human tonsil (B) and Daudi cell lysate (C) samples (35µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of F3.3 (Ab00185-23.0) 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 (c.f. Klitz et al, PMID: 12974796). Ab00185-23.0 successfully detected human HLA class II histocompatibility antigen DR in human spleen and tonsil lysate as well as Daudi cell lysate.
Flow-cytometry using the anti-HLA II DR antibody F3.3 (Ab00185) Daudi cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of F3.3 (Ab00185-23.0, 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.