Invitrogen

CD3 Monoclonal Antibody (UCHT1), NovaFluor™ Blue 585, eBioscience™

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

H002T03B04-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.4 µg)/test
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Product Specifications

Species Reactivity

Human

Published species

Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

UCHT1

Conjugate

NovaFluor™ Blue 585 NovaFluor™ Blue 585 NovaFluor™ Blue 585

Excitation/Emission Max

493/586 nm View spectra spectra

Form

Liquid

Concentration

4 µL/Test

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_3097918 (AB_2896417)

Product Specific Information

Description
The UCHT1 monoclonal antibody reacts with human CD3e, a 20 kDa subunit of the TCR complex. Along with the other CD3 subunits gamma and delta, the epsilon chain is 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 via immobilized UCHT1 initiates an intracellular biochemical pathway resulting in cellular activation and proliferation.

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

Applications Tested
This UCHT1 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at (5 µL (0.4 µg)/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: 493 nm; Emission: 586 nm; Laser: 488 nm (Blue) Laser

Target Information

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.

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.

Bioinformatics

Protein Aliases: CD247; CD247 antigen, zeta subunit; CD3 antigen, delta subunit; CD3 delta; CD3 TCR complex; CD3-epsilon; CD3d; CD3d antigen, delta polypeptide (TiT3 complex); CD3d molecule, delta (CD3-TCR complex); CD3e; CD3e antigen, epsilon polypeptide (TiT3 complex); CD3e molecule, epsilon (CD3-TCR complex); CD3g; CD3g antigen, gamma polypeptide (TiT3 complex); CD3g molecule, epsilon (CD3-TCR complex); CD3g molecule, gamma (CD3-TCR complex); CD3Z antigen, zeta polypeptide (TiT3 complex); CD3zeta chain; FLJ18683; Leu-4; OKT3, delta chain; T cell antigen receptor complex epsilon subunit of T3; 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 T3 delta chain; T-cell receptor T3 gamma chain; T-cell receptor T3 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; T3/TCR complex; TCR zeta chain

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Gene Aliases: CD247; CD3-DELTA; CD3-GAMMA; CD3-ZETA; CD3D; CD3E; CD3G; CD3H; CD3Q; CD3Z; IMD17; IMD18; IMD19; IMD25; T3D; T3E; T3G; T3Z; TCRE; TCRZ

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UniProt ID: (Human) P20963, (Human) P04234, (Human) P07766, (Human) P09693

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Entrez Gene ID: (Human) 919, (Human) 915, (Human) 916, (Human) 917

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Function(s)
transmembrane signaling receptor activity protein binding identical protein binding protein homodimerization activity transcription coactivator activity protein heterodimerization activity receptor signaling protein activity SH3 domain binding protein kinase binding receptor signaling complex scaffold activity T cell receptor binding
Process(es)
positive regulation of T cell anergy protein complex assembly cell surface receptor signaling pathway transmembrane receptor protein tyrosine kinase signaling pathway signal complex assembly G-protein coupled receptor signaling pathway response to nutrient positive regulation of gene expression negative regulation of gene expression T cell costimulation positive regulation of interferon-gamma production positive regulation of interleukin-4 production positive regulation of T cell proliferation T cell activation regulation of apoptotic process negative thymic T cell selection positive regulation of interleukin-2 biosynthetic process negative regulation of smoothened signaling pathway positive regulation of alpha-beta T cell proliferation positive regulation of peptidyl-tyrosine phosphorylation regulation of immune response positive regulation of calcium-mediated signaling T cell receptor signaling pathway apoptotic signaling pathway T cell differentiation positive thymic T cell selection positive regulation of transcription from RNA polymerase II promoter viral process Fc-gamma receptor signaling pathway involved in phagocytosis regulation of defense response to virus by virus establishment or maintenance of cell polarity protein transport regulation of lymphocyte apoptotic process
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