Anti-Fc?RII (human) Antibody, clone AT10

Application

Flow Cytometry Analysis: 25 µg/mL from a representative lot was conjugated with FITC and detected FcγRII surface expression among the monocytes, but not the lymphocytes population of human peripheral blood mononuclear cells (PBMCs) by flow cytometry (Courtesy of Professor Martin J. Glennie, University of Southampton, UK).
Flow Cytometry Analysis: A representative lot was conjugated with Phycoerythrin (PE) and immunostained the surface of human Burkitt′s lymphoma Ramos cells transfected with FcγRIIB, but not untransfected Ramos cells (Courtesy of Professor Martin J. Glennie, University of Southampton, UK).
Flow Cytometry Analysis: Clone AT10 hybridoma culture supernatant was employed to detect FcγRII-positive peripheral blood lymphocytes (PBLs) by flow cytometry (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).
Immunoprecipitation Analysis: A representative lot immunoprecipitated FcγRII from human K562 erythroleukemic cells (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).
Affinity Binding Assay: Affinity binding study using the Fab′ fragment of clone AT10 showed an equilibrium binding constant (Ka) of 5.3 x 10^8/M and a total of 1.5 x 10^5 binding sites per K562 cell (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).
Neutralization Analysis: The F(ab′)2 fragment of clone AT10 blocked FcγRII-dependent B-cell activation by a chimeric anti-CD40 mAb with human IgG1 Fc (ChiLob 7/4 h1) induced in the presence of FcγRII-overexpressing 293F as the crosslinking cells (White, A.L., et al. (2015). Cancer Cell 27(1):138–148).
Neutralization Analysis: Both the Fab′ and F(ab′)2 fragments of clone AT10, but not control IgG1 or control F(ab′)2, blocked K562 cells from rosetting with rabbit IgG-coated chick red blood cells (CRBCs) (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).
Neutralization Analysis: The F(ab′)2 fragment of clone AT10 blocked the lysis of chick red blood cells (CRBCs) by effector cells via redirected cellular cytotoxicity (RCC; antibody-dependent cell-mediated cytolysis; ADCC) mediated by an anti-CRBC monoclonal antibody (E11C12) (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).

Research Sub CategoryImmunoglobulins & Immunology

Anti-FcγRII (human) Antibody, clone AT10 is an antibody against FcγRII for use in Flow Cytometry.

Research CategoryInflammation & Immunology

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

FcγRI/CD64, FcγRII/CD32, FcγRIII/CD16, and FcγRIV/CD16-2 represent the four known classes of IgG Fc receptors (FcγRs). FcγRII, FcγRIII, and FcγRIV are low-affinity receptors for monomeric IgG, whereas FcγRI is the only high-affinity FcγR. FcγRs play important roles in inflammatory cell activation, clearance, presentation of Ag, and maintenance of IgG homeostasis. In addition to binding immune complex (IC), FcγRs have been shown to bind non-IgG ligands. For example, FcγRII is shown to bind oxLDL, while FcγRIII binding to an E. coli component is reported to negatively regulate the function of macrophage receptor with collagenous structure (MARCO). FcyRII activation is also reported to transduce a negative regulatory signal against CD38 crosslinking-induced proliferation of resting mature B cells. Low affinity immunoglobulin gamma Fc region receptor II-a/b/c (IgG Fc receptor II-a/-b/-c, CDw32, Fc-gamma RII-a/-b/-c, Fc-gamma-RIIa/b/c, FcRII-a/-b/-c, CD32) are encoded by the FCGR2A/B/C (CD32, FCG2, IGFR2) genes in human (UniProt P12318, P31994, P31995; Gene ID 2212, 2213, 9103). The FcγRIIs encoded by the three genes are highly conserved in their extracellular portion, with significant variation in the transmembrane and intracellular regions. All three types of transcripts alternative splicing to generate additional isoforms.

Immunogen

Human erythroleukaemic K562 cells.

Epitope: Extracellular domain.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in PBS without preservatives.

Format: Purified

Quality

Evaluated by Flow Cytometry analysis of human PBMCs.

Flow Cytometry Analysis: 1.0 µg of this antibody detected FcγRII in 1x10E6 human peripheral blood mononuclear cells (PBMCs).

Specificity

Clone AT10 targets the extracellular domain of human FcγRII and blocks its Fc binding activity. Expected to bind all three human FCGR2 genes-encoded FcγRII spliced isoforms reported by UniProt (P12318, P31994 & P31995).

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

31.30 kDa (FcγRIIA), 29.49 kDa (FcγRIIB1), 27.39 kDa (FcγRIIB2), 29.22 kDa (FcγRIIB3), 31.03 kDa (FcγRIIC1), 25.69 kDa (FcγRIIC2), 24.82 kDa (FcγRIIC3), 21.35 kDa (FcγRIIC4) calculated. ~42 kDa (glycosylated) and ~36 kDa (PNGase F treated) reported (Greenman, J., et al. (1991). Mol. Immunol. 28(11):1243-1254).