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CD3 antibody | MEM-57

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MCA2184F
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Mouse anti Human CD3 (MEM-57) is a monoclonal antibody recognizing an extracellular epitope of the human CD3 complex containing either δ-ε or γ-ε subunit complexes (Dave et al. 1997). Mouse anti Human CD3 (MEM-57) recognizes a conformational CD3 epitope minimally requiring the presence of the CD3ε subunit extracellular domain (Transy et al. 1989) as it will recognize murine 3D054.8 hybridoma cells transfected with cDNA to express the human CDε subunit lacking its cytoplasmic domain. CD3 serves as a pan T cell marker, it is expressed on the surface of over 95% of circulating human peripheral T cells. CD3 is also present on ~60-80% of thymocytes and on Purkinji cells, neurons present in the cerebellar cortex. CD3 is not expressed on B cells or NK cells.

The TCR complex consists of TCR heterodimers, either α/β or γ/δ expressed in a mutually exclusive manner, analagous that seen in mice on different T cell lineages. In order for the TCR heterodimer to be expressed on the cell surface it must associate with a minimal CD3 complex. The CD3 complex itself is made up of four subunits, γ, δ, ε and ζ responsible for effective signalling upon TCR activation, via a single or multiple, depending on the subunit, immunoreceptor tyrosine-based activation motif (ITAM) present in the CD3 subunit cytoplasmic domain (Kuhns et al. 2006, Dave et al. 1997a, Dave et al. 1997b).

Mouse anti Human CD3 clone MEM-57 demonstrates a mitogenic effect on peripheral blood T cells and on the lymphoblastoid Jurkat cell line (Batista et al. 2004 Brdicková et al. 2003). Clone MEM-57 immunoprecipitates proteins of ~22 kDa and more weakly ~28 kDa. from peripheral blood T cell lysates.

Target Species
Human
Species Cross-Reactivity
Target SpeciesCross Reactivity
Chimpanzee
N.B. Antibody reactivity and working conditions may vary between species.
Product Form
Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
Preparation
Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.09% sodium azide (NaN3)
1% bovine serum albumin
Approx. Protein Concentrations
IgG concentration 0.1 mg/ml
Fusion Partners
Spleen cells from immunised BALB/c mice were fused with cells of the P3/NS1/1-Ag4-1 mouse myeloma cell line.
Max Ex/Em
Fluorophore Excitation Max (nm) Emission Max (nm)
FITC 490 525
Regulatory
For research purposes only
Guarantee
12 months from date of despatch

This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry Neat
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Flow Cytometry
Use 10μl of the suggested working dilution to label 106 cells in 100μl or 100μl whole blood

Description Product Code Applications Pack Size List Price Your Price Quantity
Mouse IgG2a Negative Control:FITC MCA929F F 100 Tests loader
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Description Mouse IgG2a Negative Control:FITC

Description Product Code Applications Pack Size List Price Your Price Quantity
Human Seroblock BUF070A F 50 Test
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Description Human Seroblock
Human Seroblock BUF070B F 200 Test
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Description Human Seroblock

References for CD3 antibody

  1. Horejsí, V. et al. (1988) Monoclonal antibodies against human leucocyte antigens. II. Antibodies against CD45 (T200), CD3 (T3), CD43, CD10 (CALLA), transferrin receptor (T9), a novel broadly expressed 18-kDa antigen (MEM-43) and a novel antigen of restricted expression (MEM-74).
    Folia Biol (Praha). 34 (1): 23-34.
  2. Transy, C. et al. (1989) Most anti-human CD3 monoclonal antibodies are directed to the CD3 epsilon subunit.
    Eur J Immunol. 19: 947-50.
  3. Amlot, P.L. et al. (1996) Activation antigen expression on human T cells. I. Analysis by two-colour flow cytometry of umbilical cord blood, adult blood and lymphoid tissue.
    Clin Exp Immunol. 105: 176-82.
  4. Dave, V.P. et al. (1998) Altered functional responsiveness of thymocyte subsets from CD3delta-deficient mice to TCR-CD3 engagement.
    Int Immunol. 10: 1481-90.
  5. Morris, R.J. et al. (2005) A high-efficiency system of natural killer cell cloning.
    J Immunol Methods. 307: 24-33.
  6. Gunnlaugsdottir, B. et al. (2008) Naive human T-cells become non-responsive towards anti-TNFalpha (infliximab) treatment in vitro if co-stimulated through CD28.
    Scand J Immunol. 68: 624-34.
  7. Bughani U et al. (2017) T cell activation and differentiation is modulated by a CD6 domain 1 antibody Itolizumab.
    PLoS One. 12 (7): e0180088.

Flow Cytometry

RRID
AB_324176
UniProt
P04234
P07766
P09693
Entrez Gene
CD3D
CD3E
CD3G
GO Terms
GO:0016021 integral to membrane
GO:0004888 transmembrane receptor activity
GO:0005737 cytoplasm
GO:0031295 T cell costimulation
GO:0045059 positive thymic T cell selection
GO:0046982 protein heterodimerization activity
GO:0050852 T cell receptor signaling pathway
GO:0005057 receptor signaling protein activity
GO:0005887 integral to plasma membrane
GO:0007172 signal complex assembly
GO:0007186 G-protein coupled receptor protein signaling pathway
GO:0009897 external side of plasma membrane
GO:0017124 SH3 domain binding
GO:0019901 protein kinase binding
GO:0030159 receptor signaling complex scaffold activity
GO:0042608 T cell receptor binding
GO:0006461 protein complex assembly
GO:0007163 establishment or maintenance of cell polarity
GO:0015031 protein transport
GO:0042981 regulation of apoptosis
GO:0042110 T cell activation

MCA2184F

1013R 165250

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