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Application
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CD3e Antibody (M002T02Y06-A) in Flow
C57BL/6 mouse splenocytes were either left unstained (blue histogram) or stained with 1 µg of CD3 Monoclonal Antibody, NovaFluor Yellow 700 (purple histogram) and acquired in the YG7 channel on a 5-laser Cytek Aurora. Cells in the lymphocyte gate were used in the analysis.
Product Details
M002T02Y06-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Armenian hamster
/ IgG
Class
Monoclonal
Type
Antibody
Clone
145-2C11
Conjugate
Excitation/Emission Max
551/701 nm
View 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_3098900 (AB_2896719)
Product Specific Information
Description
The 145-2C11 monoclonal antibody reacts with mouse 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. Binding of 145-2C11 to TCR initiates the intracellular biochemical pathway resulting in cellular activation, proliferation, and apoptosis depending on specific conditions utilized. 145-2C11 is commonly used as a phenotypic marker for mouse T cells.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 145-2c11 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may 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 (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
Our internal testing shows that NovaFluor Yellow 700 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 551 nm; Emission: 701 nm; Laser: 561 nm (Yellow) 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.
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Bioinformatics
Protein Aliases: CD247; CD247 antigen; CD3 antigen, zeta polypeptide; CD3 TCR complex; CD3-zeta/eta; CD3d; CD3e; CD3g; FLJ18683; Leu-4; T cell antigen receptor complex epsilon subunit of T3; 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 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
Gene Aliases: 4930549J05Rik; A430104F18Rik; AI504783; AW552088; Cd247; CD3; Cd3-eta; Cd3-zeta; Cd3d; Cd3e; CD3epsilon; Cd3g; Cd3h; Cd3z; Cd3zeta; Ctg-3; Ctg3; T3d; T3e; T3g; T3z; Tcrk; Tcrz
UniProt ID: (Mouse) P24161, (Mouse) P04235, (Mouse) P22646, (Mouse) P11942
Entrez Gene ID: (Mouse) 12503, (Mouse) 12500, (Mouse) 12501, (Mouse) 12502
Performance Guarantee
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