CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97]
Catalog# ET1701-57
CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97]
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WB
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IF-Cell
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IHC-P
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FC
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IP
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IF-Tissue
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Human
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unconjugated
Overview
Product Name
CD3D Recombinant Rabbit Monoclonal Antibody [JJ08-97]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human CD3D aa 60-171 / 171.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC, IP, IF-Tissue
Target Molecular Weight
Predicted band size: 19 kDa
Positive Control
Jurkat cell lysates, Jurkat, human spleen tissue, human thymus tissue, human tonsil tissue.
Conjugation
unconjugated
Clone Number
JJ08-97
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:100
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IHC-P
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1:200-1:1,000
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FC
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1:1,000
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IP
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Use at an assay dependent concentration.
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IF-Tissue
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1:1,000
Target
Function
The T cell antigen receptor (TCR) recognizes foreign antigens and translates such recognition events into intracellular signals that elicit a change in the cell from a dormant to an activated state. Much of this signaling process can be attributed to a multisubunit complex of proteins that associates directly with the TCR. This complex has been designated CD3 (cluster of differentiation 3). It is composed of five invariant polypeptide chains that associate to form three dimers: a heterodimer of gamma and epsilon chains (γε), a heterodimer of delta and epsilon chains (δε) and a homodimer of two zeta chains (ΩΩ) or a heterodimer of zeta and eta chains (Ωh). The Ω and h chains are encoded by the same gene but differ in their carboxyl-terminal ends due to an alternative splicing event. The γ, ε and δ chains each contain a single copy of a conserved immunoreceptor tyrosine-based activation motif (ITAM). In contrast, the Ω chain contains three consecutive copies of the same motif. Phosphorylated ITAMs act as docking sites for protein kinases such as ZAP-70 and Syk and are also capable of regulating their kinase activity. The crystal structure of the ZAP-70 SH2 domains bound to the Ω chain ITAMs has been solved.
Background References
1. Li X., et al. 2015. Identification of a FOXP3(+)CD3(+)CD56(+) population with immunosuppressive function in cancer tissues of human hepatocellular carcinoma. Sci Rep 5:14757-14757.
2. He Y., et al. 2015. Identification of the Docking Site for CD3 on the T Cell Receptor beta Chain by Solution NMR. J. Biol. Chem. 290:19796-19805.
Tissue Specificity
CD3D is mostly present on T-lymphocytes with its TCR-CD3 partners. Present also in fetal NK-cells.
Post-translational Modification
Phosphorylated on Tyr residues after T-cell receptor triggering by LCK in association with CD4/CD8.
Subcellular Location
Membrane.
UNIPROT
Synonyms
CD3 antigen delta subunit antibody
CD3 delta antibody
CD3d antibody
CD3d antigen delta polypeptide (TiT3 complex) antibody
CD3d molecule delta (CD3-TCR complex) antibody
CD3D_HUMAN antibody
IMD19 antibody
OKT3 delta chain antibody
T cell receptor T3 delta chain antibody
T-cell receptor T3 delta chain antibody
ExpandImages
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Western blot analysis of CD3D on Jurkat cell lysates with Rabbit anti-CD3D antibody (ET1701-57) at 1/1,000 dilution.
Lysates/proteins at 15 µg/Lane.
Predicted band size: 19 kDa
Observed band size: 23 kDa
Exposure time: 17 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-57) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of Jurkat cells labeling CD3D with Rabbit anti-CD3D antibody (ET1701-57) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD3D antibody (ET1701-57) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD3D antibody (ET1701-57) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-CD3D antibody (ET1701-57) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD3D antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-57, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Application: IF-Tissue
Species: Human
Site: spleen
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000 -
Flow cytometric analysis of Jurkat cells labeling CD3D.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-57, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"