Human Fc gamma RIII (CD16) Antibody

Catalog #: MAB43251 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB43251-100
MAB43251-SP

Detection of Fc gamma RIII (CD16) in Human PBMCs by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Fc gamma RIII (CD16) Antibody Summary

Species Reactivity

Human

Specificity

Detects human Fc gamma RIIIA (CD16a) in direct ELISAs. In direct ELISAs, 60% cross-reactivity with Fc gamma RIIIB (CD16b) is observed.

Source

Monoclonal Mouse IgG_2A Clone # 1001005

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Human embryonic kidney cell, HEK293-derived human Fc gamma RIIIA/CD16a
Gly17-Gln208
Accession # P08637

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

Immunohistochemistry

5-25 µg/mL

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of Fc gamma RIII (CD16) in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with (A) Mouse Anti-Human Fc gamma RIII (CD16) Monoclonal Antibody (Catalog # MAB43251) or (B) Mouse IgG2A isotype control antibody (Catalog # MAB003) followed by anti-Mouse IgG APC-conjugated Secondary Antibody (Catalog # F0101B) and Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (Catalog # FAB3832P). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry

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Fc gamma RIII (CD16) in Human PBMCs. Fc gamma RIII (CD16) was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human Fc gamma RIII (CD16) Monoclonal Antibody (Catalog # MAB43251) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunohistochemistry

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Fc gamma RIII (CD16) in Human Spleen Tissue. Fc gamma RIII (CD16) was detected in immersion fixed paraffin-embedded sections of human spleen tissue using Mouse Anti-Human Fc gamma RIII (CD16) Monoclonal Antibody (Catalog # MAB43251) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface and cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Fc gamma RIII (CD16)

Fc gamma RIIIa is a low/intermediate affinity receptor for polyvalent immune-complexed IgG. It is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody-dependent cytotoxicity and clearance of immune complexes (1, 2). In humans, it is a 50-70 kDa type I transmembrane activating receptor expressed by NK cells, T cells, monocytes, and macrophages (1). Fc gamma RIIIb is highly related, sharing 97% amino acid (aa) identity within the extracellular domain (ECD), but is a GPI-linked receptor expressed on human neutrophils and eosinophils (1, 2). The ECD of Fc gamma RIIIa shares 63%, 61%, 65%, 59% and 58% aa identity with mouse Fc gamma RIV, rat Fc gamma RIIIa, feline CD16, bovine CD16 and porcine Fc gamma RIIIb paralogs, respectively. The Fc gamma RIIIa cDNA encodes 254 aa including a 16 aa signal sequence, 191 aa ECD with two C2-type Ig-like domains and five potential N-glycosylation sites, a 22 aa transmembrane (TM) sequence and a 25 aa cytoplasmic domain. In humans, a single nucleotide polymorphism creates high binding (176V) and low binding (176F) forms that, when homozygous, may influence susceptibility to autoimmune diseases or response to therapeutic IgG antibodies (3, 4). Catalog # 4325-FC is expressed as the 176V isoform of Fc gamma RIIIa. Fc gamma RIIIa surface expression requires interaction of an accessory chain, either the common gamma -chain or CD3 zeta (5, 6). Glycosylation patterns, electrophoretic mobility and binding affinity appear to differ between NK cell and monocyte Fc gamma RIIIa (7). The ECD of both Fc gamma RIIIa and b can be proteolytically cleaved and retain binding activity in soluble form (8-11). In monocytes and macrophages, activation and phagocytosis can trigger Fc gamma RIIIa release (11). Soluble Fc gamma RIII can be detected in normal plasma and is increased in rheumatoid arthritis and in coronary artery diseases (9, 10).

References