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Exbio
—
Research products
—
Antibodies
—
CD and related antigens
—
Anti-Hu CD3 zeta (pY72) Purified
Anti-Hu CD3 zeta (pY72) Purified
Regulatory status
RUO
Antigen
CD3 zeta (pY72)
Clone
EM-26
Format
Purified
Reactivity
Mouse, Human
Application
FC-IC, WB, ICC
Variant
0.1 mg
11-712-C100
In stock
147.40 USD
0.025 mg
11-712-C025
Delivery 1 week
73.70 USD
Variant
0.1 mg
11-712-C100
In stock
147.40 USD
0.025 mg
11-712-C025
Delivery 1 week
73.70 USD
Contact distributor
Product details
Description
Images
References
SDS download
Isotype
Mouse IgG2b
Specificity
The mouse monoclonal antibody EM-26 recognizes phosphorylated intracellular tyrosine 72 of CD3 zeta chain (CD247), which is a component of TCR/CD3 complex expressed on T cells.
Application
FC-IC, WB, ICC
Application details
Western blotting: Recommended dilution: 2 - 5 μg/ml; positive control: Jurkat cells lysate treated with pervanadate, splenocyte lysate of Balb/c or F1 mouse treated with pervanadate, non-reducing conditions recommended.
Flow cytometry: Intracellular staining; recommended dilution: 1-9 μg/ml; positive control: Jurkat cells treated with pervanadate, T-cells from lymph nodes of OT-1 mouse treated with pervanadate.
Reactivity
Mouse, Human
Immunogen
A phospho specific peptide corresponding to the amino acids surrounding tyrosine 72 of mouse CD3 zeta linked to KLH
Concentration
1 mg/ml
Preparation
Purified by protein-A affinity chromatography.
Formulation
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Storage and handling
Store at 2-8°C. Do not freeze.
Exbio licence note
The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.
Other names
CD3 zeta chain, CD247, T3Z, TCRz, IMD25
Antigen description
CD3 complex is crucial in transducing antigen-recognition signals into the cytoplasm of T cells and in regulating the cell surface expression of the TCR complex. T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta (CD247). These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR-induced growth arrest, cell survival and proliferation.
Entrez Gene ID
919
UniProt ID
P20963
Western blotting analysis using monoclonal antibodies EM-26 (anti-CD3 zeta phospho-Tyr72) and EM-17 (anti-CD3 zeta phospho-Tyr153) reacting with phosphorylated particular human CD3 zeta mutants. The Y1F and Y6F mutatants lack phosphotyrosine 72 and 153, respectively.
Separation of pervanadate stimulated Jurkat cell suspension stained using anti-human CD3 zeta (pY72) (EM-26) purified antibody (concentration in sample 3 μg/ml, GAM APC, red-filled) from pervanadate stimulated Jurkat cell suspension unstained by primary antibody (GAM APC, black-dashed) in flow cytometry analysis (intracellular staining).
Anti-Hu CD3 zeta (p72) purified antibody (clone EM-26) works in Flow Cytometry application
Analysis of the antibody staining was performed on Jurkat cells treated or untreated with pervanadate (PV) prior to the fixation and permeabilization of cell suspension with cold methanol.
Anti-Hu CD3 zeta (pY72) purified antibody (concentration in sample 1 μg/ml, red-filled histogram) binds specifically to phosphorylated tyrosine 72 (pY72) of CD3 zeta chain in PV treated, methanol permeabilized Jurkat cells (upper panel), but not to untreated methanol permeabilized control cells (lower panel).
Level of non-specific binding was assessed using Mouse IgG1 isotype control PE (MOPC-21) under same conditions (concentration in sample 1 μg/ml, black-dashed histogram).
Immunocytochemistry detection of phosphorylated CD3 zeta (EM-26; light blue) in immunological synapse formed between the lymph node naïve T cells from AND TCR transgenic mice and DCEK cells loaded with MCC peptide, after 20 min. Total CD3 zeta indicated in green, actin cytoskeleton in red.
Anti-Hu CD3 zeta (pY72) Purified (clone EM-26) specificity verification by WB.
The specificity of EM-26 antibody to phosphorylated Tyr 72 (CD3 zeta chain) was assessed by analysis of binding signals in HEK293T transfected with CD3 zeta/ZAP-70 construct followed by pervanadate (PV) treatment in comparison to the series of control cells – PV untreated transfectants, and both PV treated and untreated mock HEK293T cells.
Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer with PhosSTOP and pervanadate), mixed and heated (100°C, 5 min) with non-reducing SDS-loading buffer. Samples were resolved using 15% SDS-PAGE gel.
Nitrocellulose membrane blot was probed with mouse IgG2b monoclonal antibody EM-26 (1 µg/ml). Subclass-specific secondary antibody IRDye 680LT Goat-anti-Mouse IgG (red) was used for fluorescent Western blot detection.
Product specific references:
Dopfer EP, Schöpf B, Louis-Dit-Sully C, Dengler E, Höhne K, Klescová A, Prouza M, Suchanek M, Reth M, Schamel WW: Analysis of novel phospho-ITAM specific antibodies in a S2 reconstitution system for TCR-CD3 signalling. Immunol Lett. 2010 May 4;130(1-2):43-50
PubMed
Borroto A, Arellano I, Dopfer EP, Prouza M, Suchànek M, Fuentes M, Orfao A, Schamel WW, Alarcón B: Nck recruitment to the TCR required for ZAP70 activation during thymic development. J Immunol. 2013 Feb 1;190(3):1103-12.
PubMed
Further SDS language mutations available for download below. Please contact us with request for additional languages on info@exbio.cz
MPAbNaN3_SDS_v1_AU.pdf
MPAbNaN3_SDS_v1_GB.pdf
MPAbNaN3_SDS_v1_TR.pdf
MPAbNaN3_SDS_v6_AT.pdf
MPAbNaN3_SDS_v6_CH.pdf
MPAbNaN3_SDS_v6_CS.pdf
MPAbNaN3_SDS_v6_EN.pdf
MPAbNaN3_SDS_v6_ES.pdf
MPAbNaN3_SDS_v6_FR.pdf
MPAbNaN3_SDS_v6_IT.pdf
MPAbNaN3_SDS_v6_NO.pdf
MPAbNaN3_SDS_v6_PL.pdf
MPAbNaN3_SDS_v6_PT.pdf
MPAbNaN3_SDS_v6_SE.pdf
MPAbNaN3_SDS_v6_SK.pdf
MPAbNaN3_SDS_v6_SL.pdf
MPAbNaN3_SDS_v7_DE.pdf
Variant
0.1 mg
11-712-C100
In stock
147.40 USD
0.025 mg
11-712-C025
Delivery 1 week
73.70 USD
Variant
0.1 mg
11-712-C100
In stock
147.40 USD
0.025 mg
11-712-C025
Delivery 1 week
73.70 USD
Contact distributor
Datasheet download
Bulk requests
Technical support for this product
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