Anti-FcγRIIb/c Antibody, clone 4F5

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.

Clone 4F5 recognizes human FcγRIIb and FcγRIIc, but not any of the two polymorphic forms of human FcγRIIa with either Arg131 or His131 (Li, X., et al. (2013). Sci. Transl. Med. 5(216):216ra175; Su, K., et al. (2007). J. Immunol. 178(5):3272-3280). Expected to bind all spliced isoforms of human FCGR2B and FCGR2C reported by UniProt (P31994 & P31995).

Epitope: Extracellular domain.

Recombinant human FcγRIIb extracellular domain.

Anti-FcγRIIb/c Antibody, clone 4F5 is an antibody against FcγRIIb/c for use in Immunohistochemistry (Paraffin), Flow Cytometry, Immunoprecipitation, Western Blotting, Immunocytochemistry.

Immunohistochemistry Analysis: A 1:50 dilution from a representative lot detected FcγRIIb/c immunoreactivity in germinal center cells in human tonsil tissue sections.
Flow Cytometry Analysis: A representative lot was conjugated with Alexa Fluor^™ 647 and detected (at 10 µg/mL) FcγRIIb/c immunoreactivity among the CD19+ B lymphocytes population in human whole blood (Courtesy of Dr. Jeffrey C. Edberg, University of Alabama at Birmingham, USA).
Flow Cytometry Analysis: A representative lot was conjugated with Alexa Fluor 488 and detected FcγRIIb/c immunoreactivity on the surface of EBV-transformed human B cells from various donors (Li, X., et al. (2013). Sci. Transl. Med. 5(216):216ra175).
Flow Cytometry Analysis: A representative lot was conjugated with Alexa Fluor 488 and employed to detect FcγRIIb expression on various human PBMC populations. Highest FcγRIIb level was found on CD19+ B lymphocytes, while lower expression was found among CD14+ monocytes/neutrophils and CD3+ T-cells were negative of FcγRIIb staining (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).
Flow Cytometry Analysis: Clone 4F5 hybridoma supernatant immunostained murine B lymphoma IIA1.6 cells expressing exogenously transfected human FcγRIIb, but not any of the two polymorphic forms of human FcγRIIa with either Arg131 or His131 (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).
Immunoprecipitation Analysis: A representative lot immunoprecipitated both FcγRIIb and FcγRIIc from EBV-transformed human B cells and primary human CD19+ cells (Li, X., et al. (2013). Sci. Transl. Med. 5(216):216ra175).
Immunoprecipitation Analysis: A representative lot immunoprecipitated exogenously expressed human FcγRIIb, but not any of two polymorphic forms of human FcγRIIa with either Arg131 or His131 from murine B lymphoma IIA1.6 transfectants expressing the respective human proteins (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).
Western Blotting Analysis: A representative lot detected exogenously expressed human FcγRIIb, but not any of two polymorphic forms of human FcγRIIa with either Arg131 or His131 in lysates from murine B lymphoma IIA1.6 transfectants (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).
Immunocytochemistry Analysis: A representative lot was conjugated with Alexa Fluor 488 and detected FcγRIIb/c immunoreactivity on the surface of EBV-transformed human B cells from various donors (Li, X., et al. (2013). Sci. Transl. Med. 5(216):216ra175).
Functional Analysis: Cross-linking by goat anti-mouse IgG (GαM) of clone 4F5 F(ab′)2-bound human FcγRIIb and endogenous surface IgG-BCR on IIA1.6 murine B lymphoma cells expressing transfected human FcγRIIb resulted in a reduced calcium response than that triggered by GαM in the absence of clone 4F5 F(ab′)2. Cross-linking of FcγRIa with clone 4F5 F(ab′)2-bound human FcγRIIb on the surface of U937 cells likewise reduced the calcium response triggered by FcγRIa cross-linking alone (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).
ELISA Analysis: Clone 4F5 hybridoma supernatant detected recombinant extracellular domain (EC) of human FcγRIIb, but not any of the two polymorphic forms of recombinant human FcγRIIa ECs with either Arg131 or His131 (Su, K., et al. (2007). J. Immunol. 178(5):3272-3280).

Research Category
Inflammation & Immunology

Research Sub Category
Immunoglobulins & Immunology

Evalulated by Immunohistochemistry in human pancreas tissue.

Immunohistochemistry Analysis: A 1:50 dilution of this antibody detected FcγRIIb/c immunoreactivity in Islets of Langerhans in human pancreas tissue sections.

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.

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in PBS without preservatives.

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.

Concentration: Please refer to lot specific datasheet.

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