Invitrogen™ CD3 Monoclonal Antibody (OKT3), Super Bright™ 600, eBioscience™, Invitrogen™

Catalog No.	Quantity
63-003-741	25 Tests
63-003-742	100 Tests

Catalog No. 63-003-741 Supplier Invitrogen™ Supplier No. 63003741

Description

Mouse Monoclonal Antibody

Description: The OKT3 monoclonal antibody reacts with an epitope on the epsilon-subunit within the human CD3 complex. The OKT3 antibody has been reported to have potent immunosuppressive properties in vivo and has been proven effective in the treatment of renal, heart and liver allograft rejection. The CD3 subunits, gamma, delta, and epsilon chains, are required for proper assembly, trafficking and surface expression of the TCR complex. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells. Crosslinking of TCR initiates an intracellular biochemical pathway resulting in cellular activation and proliferation. Antibody clones OKT3 and SK7 see different epitopes. Applications Reported: This OKT3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This OKT3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.25 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. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

The CD3 complex, composed of gamma, delta, epsilon, and zeta subunits, is essential for the assembly, trafficking, and surface expression of the T cell receptor (TCR) complex. These subunits are structurally related members of the immunoglobulin superfamily and are encoded by closely linked genes on human chromosome 11. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells, but not on B or NK cells. The CD3 subunits play a crucial role in transducing antigen-recognition signals into the cytoplasm of T cells. The cytoplasmic tails of CD3 subunits contain a double tyrosine-based motif that associates with cytoplasmic signal transduction molecules, mediating T cell activation through the TCR. Crosslinking of the TCR initiates intracellular biochemical pathways that result in cellular activation, proliferation, and potentially growth arrest and cell survival. CD3 is present on 68-82% of normal peripheral blood lymphocytes, 65-85% of thymocytes, and Purkinje cells in the cerebellum. Decreased percentages of T lymphocytes may be observed in some autoimmune diseases. Defects in the CD3 gene are associated with CD3 immunodeficiency, highlighting its importance in immune function and regulation.

Specifications

• Antigen: CD3
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: OKT3
• Concentration: 5 μL/Test
• Conjugate: Super Bright 600
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: Cd3d
• Gene Accession No.: P04234, P07766, P09693, P20963
• Gene Alias: 4930549J05Rik; A430104F18Rik; AI504783; antigen CD3D, delta polypeptide (TiT3 complex); antigen CD3E, epsilon polypeptide (TiT3 complex); antigen CD3G, gamma polypeptide; antigen CD3Z, zeta polypeptide; AW552088; Cd247; CD247 antigen; CD247 antigen, zeta subunit; Cd247 molecule; CD3; CD3 antigen delta chain; CD3 antigen delta polypeptide; CD3 antigen gamma chain; CD3 antigen, delta polypeptide; CD3 antigen, delta subunit; CD3 antigen, epsilon polypeptide; CD3 antigen, gamma polypeptide; CD3 antigen, zeta polypeptide; CD3 delta; CD3 epsilon; CD3 epsilon chain; CD3 epsilon subunit; CD3 epsilon subunit precursor; CD3 gamma-chain; CD3 glycoprotein; CD3 glycoprotein precursor; CD3 molecule delta polypeptide; CD3 molecule, delta; CD3 molecule, epsilon; CD3 molecule, epsilon polypeptide; CD3 molecule, gamma; CD3 molecule, gamma polypeptide; CD3 protein; CD3 TCR complex; CD3 type I transmembrane glycoprotein; CD3 type I transmembrane glycoprotein precursor; CD3 zeta chain; Cd3d; CD3D antigen delta; CD3D antigen, delta polypeptide (TiT3 complex); CD3d molecule; CD3d molecule, delta (CD3-TCR complex); CD3-DELTA; Cd3e; CD3E antigen, epsilon polypeptide; CD3E antigen, epsilon polypeptide (TiT3 complex); CD3e molecule; CD3e molecule, epsilon (CD3-TCR complex); CD3epsilon; CD3-epsilon; Cd3-eta; Cd3g; CD3G antigen, gamma polypeptide; CD3g antigen, gamma polypeptide (TiT3 complex); CD3g molecule; CD3g molecule, epsilon (CD3-TCR complex); CD3g molecule, gamma (CD3-TCR complex); CD3-GAMMA; Cd3h; CD3Q; Cd3z; CD3Z antigen, zeta polypeptide (TiT3 complex); Cd3zeta; Cd3-zeta; CD3zeta chain; CD3-zeta/eta; Ctg3; Ctg-3; FLJ18683; IMD17; IMD18; IMD19; IMD25; Leu-4; OKT3, delta chain; T cell antigen receptor complex epsilon subunit of T3; T3/TCR complex; T3d; T3e; T3g; T3Z; T-cell antigen receptor complex, epsilon subunit of T3; T-cell antigen receptor complex, gamma subunit of T3; T-cell antigen receptor complex, zeta subunit of CD3; T-cell receptor CD3 epsilon chain; T-cell receptor CD3 epsilon subunit; T-cell receptor CD3 subunit zeta; T-cell receptor CD3, subunit zeta; T-cell receptor T3 delta chain; T-cell receptor T3 eta chain; T-cell receptor T3 gamma chain; T-cell receptor T3 zeta chain; T-cell receptor zeta chain; T-cell surface antigen T3/Leu-4 epsilon chain; T-cell surface glycoprotein CD3 delta chain; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface glycoprotein CD3 gamma chain; T-cell surface glycoprotein CD3 zeta chain; T-cell surface protein; TcR CD3 delta-chain; TcR CD3 gamma-chain; TCR zeta chain; TCR zeta chain subunit; TCRE; Tcrk; TCRZ; TCRzeta; TiT3 complex; type I transmembrane protein; T-cell surface molecule CD3
• Gene Symbols: Cd247, Cd3d, CD3E, CD3g
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 25 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 915, 916, 917, 919
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

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.

For Research Use Only.