Recombinant Anti-CD3D antibody [EP4426] (ab109531)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP4426] to CD3D
- Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD3D antibody [EP4426]
See all CD3D primary antibodies -
Description
Rabbit monoclonal [EP4426] to CD3D -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat cell lysate. Human thymus tissue lysate. IP: Jurkat whole cell 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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP4426 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-CD3D antibody [EP4426] (ab198935)
- Alexa Fluor® 647 Anti-CD3D antibody [EP4426] (ab198937)
- Alexa Fluor® 555 Anti-CD3D antibody [EP4426] (ab208514)
- Anti-CD3D antibody [EP4426] - Low endotoxin, Azide free (ab215040)
- Anti-CD3D antibody [EP4426] - BSA and Azide free (ab226073)
- Alexa Fluor® 568 Anti-CD3D antibody [EP4426] (ab312640)
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109531 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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Flow Cyt | (1) |
Use at an assay dependent concentration.
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WB |
1/1000. Predicted molecular weight: 19 kDa.
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IP |
1/100.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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ICC/IF |
1/500 - 1/1000.
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Notes |
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Flow Cyt
Use at an assay dependent concentration. |
WB
1/1000. Predicted molecular weight: 19 kDa. |
IP
1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
ICC/IF
1/500 - 1/1000. |
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: 915 Human
- Omim: 186790 Human
- SwissProt: P04234 Human
- Unigene: 504048 Human
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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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All lanes : Anti-CD3D antibody [EP4426] (ab109531) at 1/1000 dilution (Purified)
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 19 kDa -
Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling CD3D with Purified ab109531 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD3D with Purified ab109531 at 1:50 dilution (2.6 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD3D with Purified ab109531 at 1:100 dilution (1.3 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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All lanes : Anti-CD3D antibody [EP4426] (ab109531) at 1/1000 dilution
Lane 1 : THP1 whole cell lysate (-ve control)
Lane 2 : Raji whole cell lysate (-ve control)
Lane 3 : Jurkat whole cell lysate
Lane 4 : Human Thymus tissue lysate
Lane 5 : Mouse Thymus tissue lysate
Lane 6 : Rat Thymus tissue lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab109531 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab109531 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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Purified ab109531 at 1/50 dilution (2µg) immunoprecipitating CD3D in Jurkat whole cell lysate.
Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
Lane 2 (+): ab109531 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109531 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 23 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab109531 has been referenced in 8 publications.
- Pan J et al. Papillary Thyroid Carcinoma Landscape and Its Immunological Link With Hashimoto Thyroiditis at Single-Cell Resolution. Front Cell Dev Biol 9:758339 (2021). PubMed: 34805166
- Uchibori R et al. Functional Analysis of an Inducible Promoter Driven by Activation Signals from a Chimeric Antigen Receptor. Mol Ther Oncolytics 12:16-25 (2019). PubMed: 30662937
- Santucci-Pereira J et al. Genomic signature of parity in the breast of premenopausal women. Breast Cancer Res 21:46 (2019). PubMed: 30922380
- Peng J et al. Single-cell RNA-seq highlights intra-tumoral heterogeneity and malignant progression in pancreatic ductal adenocarcinoma. Cell Res 29:725-738 (2019). PubMed: 31273297
- Alunno A et al. Insulin-Like Growth Factor Binding Protein 6 in Rheumatoid Arthritis: A Possible Novel Chemotactic Factor? Front Immunol 8:554 (2017). PubMed: 28572803
- Jin C et al. Safe engineering of CAR T cells for adoptive cell therapy of cancer using long-term episomal gene transfer. EMBO Mol Med 8:702-11 (2016). PubMed: 27189167
- Alunno A et al. Telocytes in minor salivary glands of primary Sjögren's syndrome: association with the extent of inflammation and ectopic lymphoid neogenesis. J Cell Mol Med 19:1689-96 (2015). IHC-P ; Human . PubMed: 25753463
- Liu X et al. Immune reconstitution from peripheral blood mononuclear cells inhibits lung carcinoma growth in NOD/SCID mice. Oncol Lett 8:1638-1644 (2014). PubMed: 25202383