Invitrogen

CD16/CD32 Monoclonal Antibody (93), NovaFluor™ Red 685, eBioscience™

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Datasheet
Protocols
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Datasheet
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Product Details

M021T02R02-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

1.0 µg/test
-
Product Specifications

Species Reactivity

Mouse

Host/Isotype

Rat / IgG2a, lambda

Class

Monoclonal

Type

Antibody

Clone

93

Conjugate

NovaFluor™ Red 685 NovaFluor™ Red 685 NovaFluor™ Red 685

Excitation/Emission Max

637/685 nm View spectra spectra

Form

Liquid

Concentration

0.1 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3099012

Product Specific Information

Description
The 93 monoclonal antibody reacts with an epitope shared by mouse CD16 and CD32. CD16 (Fc gamma III Receptor) and CD32 (Fc gamma II Receptor) are the low affinity receptors for the mouse IgG Fc portion and are expressed by B cells, monocyte/macrophages, NK cells, and neutrophils.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
This 93 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 1.0 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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: 637 nm; Emission: 685 nm; Laser: 633-640 nm (Red) Laser

Target Information

CD16 is a 50-65 kDa cell surface molecule that exists in two forms - a transmembranous form expressed by natural killer (NK) cells, and some T cells and a phosphatidylinositol linked form expressed by granulocytes. CD16 is a low affinity receptor for IgG (FcR III), and is an important receptor mediating ADCC (antibody dependent cell mediated cytotoxicity) by NK cells. IN comparison, CD32 is a 40 kD glycoprotein that acts as a low affinity receptor for IgG (also known as Fc gamma RII), and mediates several functions including endocytosis, activation of secretion, cytotoxicity and immunomodulation. CD32 is expressed by B cells, monocytes, granulocytes and platelets. Further, CD32 is involved in the phagocytosis of immune complexes, the regulation of antibody production by B-cells, and variations of the CD32 gene may increase vulnerability to systemic lupus erythematosus (SLE). Several transcript variants encoding different isoforms of CD32 have been found.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

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.

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Bioinformatics

Protein Aliases: CD16A; CD16B; CD32; CD32 receptor 2; CDw32; Fc gamma receptor IIB; Fc gamma receptor III; Fc gamma RIIB; Fc-gamma RII; Fc-gamma RII-b; Fc-gamma RII-c; fc-gamma RIII; Fc-gamma-RII; fc-gamma-RIIB; Fc-gamma-RIIc; Fcg receptor III; FCG2; FcgammaRIIB; FcgammaRIII; FCGR2C; FCGR3A; FCGR3B; FCGRIII; FCR-10; FcRII-b; FcRII-c; fcRIII; FCRIIIA; FGFR2B; IgG Fc receptor II beta; igG Fc receptor III; Low affinity immunoglobulin gamma Fc region receptor II; low affinity immunoglobulin gamma Fc region receptor II-b; low affinity immunoglobulin gamma Fc region receptor III; Ly-17; Lymphocyte antigen 17; RP11-5K23.1

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Gene Aliases: AI528646; CD16; CD32; F630109E10Rik; Fc[g]RII; Fcgr2; Fcgr2a; Fcgr2b; FcgRII; Fcr-2; Fcr-3; fcRII; Ly-17; Ly-m20; LyM-1

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UniProt ID: (Mouse) P08101

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Entrez Gene ID: (Mouse) 14130, (Mouse) 14131

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Function(s)
receptor signaling protein activity protein binding IgG binding immunoglobulin binding IgG receptor activity immunoglobulin receptor superfamily
Process(es)
negative regulation of type I hypersensitivity immune complex clearance negative regulation of immunoglobulin production negative regulation of inflammatory response to antigenic stimulus negative regulation of acute inflammatory response to antigenic stimulus negative regulation of humoral immune response mediated by circulating immunoglobulin receptor-mediated endocytosis phagocytosis, engulfment defense response immune response cell surface receptor signaling pathway response to bacterium immunoglobulin mediated immune response antigen processing and presentation of exogenous peptide antigen via MHC class II negative regulation of B cell proliferation endocytic recycling negative regulation of interleukin-10 production negative regulation of neutrophil apoptotic process intracellular signal transduction mast cell activation negative regulation of phagocytosis positive regulation of phagocytosis negative regulation of immune response cellular response to molecule of bacterial origin antibody-dependent cellular cytotoxicity positive regulation of type IIa hypersensitivity positive regulation of type III hypersensitivity positive regulation of type I hypersensitivity serotonin secretion phagocytosis, recognition neutrophil chemotaxis Fc-gamma receptor signaling pathway positive regulation of tumor necrosis factor biosynthetic process antigen processing and presentation of exogenous peptide antigen via MHC class I regulation of immune response
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