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Description
This 3G8 monoclonal antibody reacts with human and non-human primate CD16, which is also known as the low-affinity Fc gamma RIII. Expression of CD16 on lymphocytes and monocytes is similar in non-human primates. However, while CD16 is not found on neutrophils in macaques and baboons, this receptor is detected on these cells in sooty mangabeys. Based on cross-blocking studies 3G8 recognizes the same epitope as CB16. However, 3G8 and B73.1 antibody clones bind distinct epitopes.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 3G8 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.6 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 508 nm; Emission: 664 nm; Laser: 488 nm (Blue) Laser
CD16, known as Fc gamma receptor III, exists in two distinct isoforms: Fc gamma RIIIA and Fc gamma RIIIB. These isoforms are encoded by the genes FCGR3A and FCGR3B, respectively, which are located on chromosome 1 and share a high degree of similarity. Fc gamma RIIIA is expressed as a polypeptide-anchored form on monocytes, macrophages, and lymphocytes such as natural killer (NK) cells, but not on T or B cells. This receptor recognizes the Fc portion of immunoglobulin G (IgG) and is involved in the clearance of immune complexes from circulation. Upon binding IgG, Fc gamma RIIIA activates signal transduction pathways that result in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, cytokine release, and antigen presentation. Fc gamma RIIIB is expressed on polymorphonuclear neutrophils (PMN) as a glycosylphosphatidylinositol (GPI)-anchored form. It plays a role in similar immune functions, including cellular-mediated cytotoxicity and enhancement of virus infections. Mutations in the FCGR3A gene have been associated with Immunodeficiency 20 and linked to susceptibility to recurrent viral infections, systemic lupus erythematosus, and alloimmune neonatal neutropenia. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. Diseases associated with FCGR3A include Immunodeficiency 20 and Herpes Zoster.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: CD16-I; CD16-II; CD16a; CD16a antigen; CD16b; CD32; CD32 receptor 2; CDw32; Fc fragment of IgG low affinity IIIa receptor; Fc fragment of IgG, low affinity III, receptor for (CD16); Fc fragment of IgG, low affinity IIIa, receptor (CD16a); Fc fragment of IgG, low affinity IIIb, receptor (CD16b); Fc gamma receptor III-A; Fc gamma receptor IIIa; Fc gamma receptor IIIb; Fc gamma RIIIa; Fc-gamma receptor III-2 (CD 16); Fc-gamma receptor IIIb (CD 16); Fc-gamma receptor IIIb (CD16); Fc-gamma RII-b; Fc-gamma RII-c; Fc-gamma RIII; Fc-gamma RIII-alpha; Fc-gamma RIII-beta; Fc-gamma RIIIa; fc-gamma RIIIb; Fc-gamma-RIIb; Fc-gamma-RIIc; FCG2; FcgammaRIIIA; FCGR2; FCGR2A; FCGR2B; FCGR2C; FcR-10; FcRII-b; FcRII-c; IgG Fc receptor III; IgG Fc receptor III-1; IgG Fc receptor III-2; IgG Fc receptor III-A; immunoglobulin G Fc receptor III; low affinity immunoglobulin gamma Fc region receptor III; Low affinity immunoglobulin gamma Fc region receptor III-A; Low affinity immunoglobulin gamma Fc region receptor III-B; neutrophil-specific antigen NA; RP11-5K23.1
Gene Aliases: CD16; CD16A; CD16B; FCG3; FCGR3; FCGR3A; FCGR3B; FCGRIII; FCR-10; FCRIII; FCRIIIA; FCRIIIb; IGFR3; IMD20
UniProt ID: (Rhesus monkey) A3RFZ7, (Human) P08637, (Human) O75015
Entrez Gene ID: (Rhesus monkey) 720006, (Human) 2214, (Human) 2215
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