Human Fc gamma  RIIIA/CD16a Antibody

Catalog # Availability Size / Price Qty
MAB4325-100
MAB4325-SP
Detection  of Human Fc gamma  RIIIA/CD16a by Western Blot.
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Human Fc gamma  RIIIA/CD16a Antibody Summary

Species Reactivity
Human
Specificity
Detects human Fc gamma RIIIA/CD16a in direct ELISAs.
Source
Monoclonal Mouse IgG2A Clone # 1001049
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 as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
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
8-25 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.

Data Examples

Western Blot Detection  of Human Fc gamma  RIIIA/CD16a by Western Blot. View Larger

Detection of Human Fc gamma  RIIIA/CD16a by Western Blot. Western blot shows lysates of human lung tissue and human peripheral blood mononuclear cells (PBMCs). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Fc gamma  RIIIA/CD16a Monoclonal Antibody (Catalog # MAB4325) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Fc gamma  RIIIA/CD16a at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry Detection of Fc gamma  RIIIA/CD16a in Human PBMCs by Flow Cytometry. View Larger

Detection of Fc gamma  RIIIA/CD16a in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs)were stained with (A) Mouse Anti-Human Fc gamma  RIIIA/CD16a Monoclonal Antibody (Catalog # MAB4325) or (B) Mouse IgG2A isotype control antibody (Catalog # MAB003) followed by Goat 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 Fc gamma  RIIIA/CD16a  in Human PBMCs. View Larger

Fc gamma  RIIIA/CD16a in Human PBMCs. Fc gamma  RIIIA/CD16a was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human Fc gamma  RIIIA/CD16a Monoclonal Antibody (Catalog # MAB4325) 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 surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Immunohistochemistry Fc gamma  RIIIA/CD16a  in Human Spleen. View Larger

Fc gamma  RIIIA/CD16a in Human Spleen. Fc gamma  RIIIA/CD16a was detected in immersion fixed paraffin-embedded sections of human spleen using Mouse Anti-Human Fc gamma  RIIIA/CD16a Monoclonal Antibody (Catalog # MAB4325) 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 cytoplasm and cell surfaces in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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 RIIIA/CD16a

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
  1. Nimmerjahn, F. and J.V. Ravetch (2006) Immunity 24:19. 
  2. Ravetch, J.V. and B. Perussia (1989) J. Exp. Med. 170:481. 
  3. Wu, J. et al. (1997) J. Clin. Invest. 100:1059. 
  4. Dall’Ozzo, S. et al. (2004) Cancer Res. 64:4664. 
  5. Kim, M.-K. et al. (2003) Blood 101:4479. 
  6. Lanier, L.L. et al. (1989) Nature 342:803.
  7. Edberg, J.C. and R.P. Kimberley (1997) J. Immunol. 159:3849.
  8. Li, P. et al. (2007) J. Biol. Chem. 282:6210.
  9. Masuda, M. et al. (2003) J. Rheumatol. 30:1911.
  10. Masuda, M. et al. (2006) Atherosclerosis 188:377.
  11. Webster, N.L. et al. (2006) J. Leukoc. Biol. 79:294.
Long Name
Fc gamma Receptor III A
Entrez Gene IDs
2214 (Human); 304966 (Rat)
Alternate Names
CD16a antigen; CD16a; CD16FCRIIIA; Fc fragment of IgG, low affinity IIIa, receptor (CD16a); Fc fragment of IgG, low affinity IIIa, receptor for (CD16); Fc gamma receptor III-A; Fc gamma RIIIA; FCG3; Fc-gamma receptor III-2 (CD 16); Fc-gamma receptor IIIb (CD16); Fc-gamma RIII; Fc-gamma RIIIa; Fc-gamma RIII-alpha; FCGR3; FCGR3A; FCGRIII; FcgRIIIA; FcR-10; FCRIII; FcRIIIa; IGFR3FcgammaRIIIA; immunoglobulin G Fc receptor III; low affinity III, receptor for (CD16); low affinity immunoglobulin gamma Fc region receptor III-A; neutrophil-specific antigen NA

Product Datasheets

FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Mouse IgG2A Isotype Control

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Reconstitution Buffers

Secondary Antibodies

Mouse IgG2A Antibody

Mouse IgG2A Antibody

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