United Kingdom (UK)
Recombinant monoclonal antibody to CD32. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma IV.3.
UniProt Accession Number of Target Protein: P12318
Alternative Name(s) of Target: CDw32; Fc-gamma RII-a; FcγRIIa; FcγRII; FcRII-a; FCGR2A; IgG Fc receptor II-a; Low affinity immunoglobulin gamma Fc region receptor II-a
Immunogen: The original antibody was generated against the FcR of K562 cells.
Specificity: This antibody recognizes and binds CD32a.
Application Notes: This antibody successfully precipitated a 40-kd FcR from lysates of U937 and K562 cells but not from Daudi or Raji cells. It selectively inhibited the binding of murine IgG1-coated erythrocytes to U937 cells. IV.3 blocked mIgG1 anti-T3 (Leu-4) stimulation but not mIgG2a anti-T3 (OKT3) stimulation of T cells (PMID: 2936805). Anti-FcyRII antibody IV.3 specifically stained Ltk- cells transfected with the FcyR cDNA clone in an immunofluorescence assay (PMID: 2824655). It was reported that IV.3 induces anaphylaxis in immunocompetent CD32a-transgenic “FCGR2A” mice and that IV.3 also induce severe thrombocytopenia in FCGR2A mice (PMID: 26396093). IV.3, reacted with CD32A-R131 exclusively, confirming the specificity in an ELISA (PMID:17386079).
Antibody first published in:
Looney et al. Human monocytes and U937 cells bear two distinct Fc receptors for IgG. J Immunol. (1986); 136(5):1641-7. PMID:2936805
Note on publication:
Describes the fact that U937 cells contain a 40kd FcR which is defined by a monoclonal antibody IV.3.
Immunofluorescence staining of U937 cells with anti-CD32 (Ab02071) IV.3 Immunofluorescence analysis of paraformaldehyde fixed U937 cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of IV.3 (Ab02071-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab02071-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-CD32 antibody IV.3 (Ab02071). Paraformaldehyde fixed U937 cells permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of IV.3 (Ab02071-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.