Recombinant Anti-CD3D antibody [EPR20544] (ab213362)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20544] to CD3D
- Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, IP
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CD3D antibody [EPR20544]
See all CD3D primary antibodies -
Description
Rabbit monoclonal [EPR20544] to CD3D -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse thymus and lymph node lysates. IHC-P: Mouse spleen and thymus tissues. IHC-Fr: Mouse spleen tissue. Flow Cyt: Mouse spleen cells. IP: Mouse thymus lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20544 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213362 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr |
1/100.
Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
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Flow Cyt |
1/600.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
1/100. Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
Flow Cyt
1/600. |
IP
1/30. |
Target
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Function
The CD3 complex mediates signal transduction. -
Involvement in disease
Defects in CD3D are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)/B(+)/NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. -
Sequence similarities
Contains 1 ITAM domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 12500 Mouse
- SwissProt: P04235 Mouse
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Form
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Alternative names
- CD3 antigen delta subunit antibody
- CD3 delta antibody
- CD3-delta antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3D with ab213362 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic staining on T cells of mouse splenic periarterial lymphatic sheath is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD3D with ab213362 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Cytoplasmic and membranous staining on mouse splenic periarterial lymphatic sheath is observed.
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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All lanes : Anti-CD3D antibody [EPR20544] (ab213362) at 1/1000 dilution
Lane 1 : Mouse thymus tissue lysate
Lane 2 : Mouse lymph node tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 19 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 15 seconds; Lane 2: 3 minutes.
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Flow cytometric analysis of mouse spleen cells labeling CD3D with ab213362 at 1/600 dilution (right panel), compared with a rabbit monoclonal IgG isotype control (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Firstly, cells surface was stained with anti-mouse CD3 Alexa Fluor 647 (Y axis), then cells were fixed with 4% PFA followed by intracellular staining with ab213362. The same population of cells were stained by both: anti-mouse CD3 and anti-mouse CD3D (ab213362) antibodies.
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CD3D was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab213362 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab213362 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: Mouse thymus lysate, 10 µg (Input).
Lane 2: ab213362 IP in mouse thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213362 in mouse thymus lysate .
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling CD3D with ab213362 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and membranous staining on T cells of mouse thymus medulla is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab213362 has not yet been referenced specifically in any publications.