Invitrogen™ CD16 Monoclonal Antibody (3G8), Super Bright™ 436, eBioscience™, Invitrogen™

Catalog No.	Quantity
62-016-642	100 Tests

Catalog No. 62-016-642 Supplier Invitrogen™ Supplier No. 62016642

Description

Mouse Monoclonal Antibody

Description: This 3G8 monoclonal antibody reacts with human and non-human primate CD16, which is also known as the low-affinity Fc gamma RIII. CD16 exists as two distinct isoforms, Fc gamma RIIIA and Fc gamma RIIIB. In humans, Fc gamma RIIIA is expressed as a polypeptide-anchored form on monocytes, macrophages, and lymphocytes such as NK cells. T and B cells do not express this Fc receptor. Fc gamma RIIIB is also detected on neutrophils as a GPI-anchored form. 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. Binding of IgG leads to activation of signal transduction pathways, resulting in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, cytokine release, and antigen presentation. Based on cross-blocking studies 3G8 recognizes the same epitope as CB16. However, 3G8 and B73.1 antibody clones bind distinct epitopes. Applications Reported: This 3G8 antibody has been reported for use in flow cytometric analysis. 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 (0.06 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.

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.

Specifications

• Antigen: CD16
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: 3G8
• Concentration: 5 μL/Test
• Conjugate: Super Bright 436
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: FCGR3B
• Gene Accession No.: A3RFZ7, O75015, P08637
• Gene Alias: 4833442P21Rik; CD16; CD16-2; CD16A; CD16a antigen; CD16b; CD32; CD32 receptor 2; CDw32; cytolytic trigger molecule G7; Fc fragment of IgG intermediate affinity IV receptor; Fc fragment of IgG low affinity IIIa receptor; Fc fragment of IgG receptor IIIa; Fc fragment of IgG receptor IIIb; Fc fragment of IgG, low affinity III, receptor for (CD16); Fc fragment of IgG, low affinity IIIa, receptor; Fc fragment of IgG, low affinity IIIa, receptor (CD16a); Fc fragment of IgG, low affinity IIIa, receptor for (CD16); Fc fragment of IgG, low affinity IIIa, receptor for (CD16); Fc fragment of IgG, low affinity III, receptor for (CD16); Fc fragment of IgG, low affinity IIIb, receptor (CD16b); Fc gamma receptor III; Fc gamma receptor IIIa; Fc gamma receptor III-A; Fc gamma receptor IIIb; Fc gamma RIIIa; Fc gammaRIV; Fc receptor, IgG, low affinity III; Fc receptor, IgG, low affinity IV; Fc receptor-like 3; Fcg receptor III; FCG2; FCG3; 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 RIIIa; Fc-gamma RIII-alpha; fc-gamma RIIIb; fc-gamma RIII-beta; Fc-gamma-RIIb; Fc-gamma-RIIc; FcgammaRIII; FcgammaRIII a.1; FcgammaRIII a.2; FcgammaRIII a.3; FcgammaRIIIA; FcgammaRIV; FCGR2; FCGR2A; FCGR2B; FCGR2C; FCGR3; Fcgr3a; FCGR3B; Fcgr4; FCGRIII; FcgRIV; FcR-10; FcRII-b; FcRII-c; FcRIII; FCRIIIA; FCRIIIb; Fcrl3; IGFR3; IgG Fc gamma receptor III; igG Fc receptor III; IgG Fc receptor III-1; igG Fc receptor III-2; IMD20; immunoglobulin G Fc receptor III; LOC100911825; 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; low affinity immunoglobulin gamma Fc region receptor III-like; Low affinity immunoglobulin gamma Fc region receptor IV; neutrophil-specific antigen NA; RP11-5K23.1; transmembrane receptor FcgammaRIII-X
• Gene Symbols: Fcgr3, FCGR3A, FCGR3B
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 2214, 2215, 720006
• Target Species: Baboon, Chimpanzee, Cynomolgus Monkey, Human, Monkey, Rhesus Macaque
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1

Frequently Asked Questions (FAQs)

Can I use the OneComp and UltraComp eBeads microspheres with Super Bright-conjugated antibodies?

UltraComp eBeads microspheres (Cat. No. 01-2222) are recommended for use with Super Bright dyes.
Note: Super Bright Staining Buffer (Cat. No. SB-4400) is not compatible with UltraComp eBeads microspheres (Cat. No. 01-2222-41, 00-2222-42). If using UltraComp eBeads microspheres as a compensation tool, solely use Flow Cytometry Stain Buffer (Cat. No. 00-4222-26, 00-4222-57) for any antibody dilutions.

In some experiments, we have observed that compensation values for Super Bright 780- and Brilliant Violet 785- or Brilliant Violet 786-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads, both with Super Bright 780 and Brilliant Violet 786. We have not tested Brilliant Violet 785 with the AbC beads.

Can I prepare an antibody cocktail containing Super Bright Staining Buffer and Super Bright-conjugated antibodies ahead of time?

We recommend that the antibody cocktails containing Super Bright-conjugated antibodies and Super Bright Staining Buffer are prepared fresh prior to staining. Discard any unused portions. We do not recommend overnight storage of prepared cocktails.

Can I fix my cells after staining with Super Bright-conjugated antibodies? How long can fixed cells be stored prior to analysis?

Samples that have been stained with antibodies conjugated to Super Bright dyes may be stored for up to three days, at 2-8°C, in the dark, using either IC Fixation Buffer (Cat. No. 00-8222) or 1-step Fix/Lyse Buffer (Cat. No. 00-5333) with no significant effect on brightness or compensation.

Are the Super Bright dyes sensitive to methanol fixation?

Super Bright dyes are stable in methanol-based fixation buffers.

Can Super Bright-conjugated antibodies be used in combination with intracellular (IC Fixation/Permeabilization) or intranuclear (Foxp3 fixation/permeabilization) staining?

Yes, Super Bright-conjugated antibodies are stable in formaldehyde-based fixation buffers and permeabilization buffers, such as the IC Fixation and Permeabilization Buffer Set (Cat. No. 88-8824) and the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523).

Which laser do I use to detect Super Bright-conjugated antibodies?

The violet laser (405 nm) should be used to excite Super Bright-conjugated antibodies.

Can the Super Bright Staining Buffer be used with other polymer dyes?

Yes, the Super Bright Staining Buffer (Cat. No. SB-4400) is compatible with other polymer dyes (i.e., Brilliant Violet dyes) and is useful for minimizing any non-specific polymer interactions when two or more of these dyes are used in combination.

Can Super Bright-conjugated antibodies be used in combination with other polymer dyes? What buffer should I use when using more than one Super Bright or polymer dye?

When using two or more Super Bright dyes, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Are there recommended tubes for staining with Super Bright-conjugated antibodies?

We recommend using polystyrene tubes (for example, FACS tubes) for staining with your Super Bright-conjugated antibodies. If using polypropylene (for example, Eppendorf tubes), protecting from light is critical.

Is there a specific buffer I should use when staining with Super Bright-conjugated antibodies?

No special buffer is required when using a single Super Bright-conjugated antibody in a panel.
When using more than one Super Bright dye, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

How does the protocol for staining with Super Bright-conjugated antibodies differ from staining with conventional organic fluorochrome and eVolve conjugated antibodies?

Antibodies conjugated to Super Bright dyes can be used similarly to traditional fluorochromes. If multiple Super Bright dyes are used in combination with each other or in combination with other polymer dyes (i.e., Brilliant Violet dyes), then the use of Super Bright Staining Buffer (Cat. No. SB-4400) is recommended to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Can the Super Bright dyes be used for non-flow applications?

Super Bright dyes have not been tested for applications other than flow cytometry.

What is the difference between the Super Bright and eVolve dyes?

Super Bright dyes are fluorochromes based on polymer technology and are excited by the violet laser. In contrast, eVolve dyes are based on Qdot nanocrystal technology and are maximally excited by the UV laser, although they may also be excited by violet, blue, yellow-green, and, depending on the eVolve, the red laser lines. The general shape of the emission spectra for Super Bright dyes will be more similar to traditional fluorochromes, while eVolve dyes have extremely narrow emission spectra. Please contact Tech Support (techsupport@thermofisher.com) for more information.

What are the Super Bright dyes?

eBioscience Super Bright dyes are a series of patent-pending fluorochromes that are based upon a fluorescent polymer and its tandems. Super Bright dyes have been developed for use in flow cytometry and can be excited by the violet laser (405 nm). The peak emission of each Super Bright dye is indicated by the number in the product description, e.g., "Super Bright 600" has a peak emission of 600 nm.

Are the Super Bright Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.