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Immunology Adaptive Immunity T Cells CD
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RecombinantRabMAb

Recombinant Anti-CD3D antibody [EP4426] (ab109531)

  • Datasheet
Reviews (1)Q&A (1)References (6)

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Western blot - Anti-CD3D antibody [EP4426] (ab109531)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD3D antibody [EP4426] (ab109531)
  • Immunoprecipitation - Anti-CD3D antibody [EP4426] (ab109531)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
  • Western blot - Anti-CD3D antibody [EP4426] (ab109531)
  • 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: WB, IP, IHC-P, ICC/IF
  • Reacts with: Human

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Overview

  • Product name

    Anti-CD3D antibody [EP4426]
    See all CD3D primary antibodies
  • Description

    Rabbit monoclonal [EP4426] to CD3D
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD3D aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P04234

  • Positive control

    • WB: Jurkat and HuT 78 cell lysates and Human thymus tissue lysates. IHC-P: Human tonsil tissue. ICC/IF: Jurkat cells. IP: Jurkat whole cell lysate.
  • 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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • 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: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP4426
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Hematopoietic Stem Cells
    • Human Lineage Negative
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • 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)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat membrane extract lysate (ab30130)
  • Recombinant Protein

    • Recombinant Human CD3D protein (ab167701)

Applications

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
WB 1/1000 - 1/10000. Predicted molecular weight: 19 kDa.
IP 1/10 - 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

  • 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.
  • Target information above from: UniProt accession P04234 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 915 Human
    • Omim: 186790 Human
    • SwissProt: P04234 Human
    • Unigene: 504048 Human
    • Form

      .
    • Alternative names

      • CD3 antigen delta subunit antibody
      • CD3 delta antibody
      • CD3-delta antibody
      • CD3d antibody
      • CD3d antigen delta polypeptide (TiT3 complex) antibody
      • CD3d molecule 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
      • T-cell surface glycoprotein CD3 delta chain antibody
      • T3D antibody
      see all

    Images

    • Western blot - Anti-CD3D antibody [EP4426] (ab109531)
      Western blot - Anti-CD3D antibody [EP4426] (ab109531)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)

      IHC image of CD3D staining in a formalin fixed, paraffin embedded human B-Cell lymphoma tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab109531, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunocytochemistry/ Immunofluorescence - Anti-CD3D antibody [EP4426] (ab109531)
      Immunocytochemistry/ Immunofluorescence - Anti-CD3D antibody [EP4426] (ab109531)

      Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CD3D with purified ab109531 at 1/500 dilution (4.5 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. 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. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.

    • Immunoprecipitation - Anti-CD3D antibody [EP4426] (ab109531)
      Immunoprecipitation - Anti-CD3D antibody [EP4426] (ab109531)

      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)

      IHC image of CD3D staining in a formalin fixed, paraffin embedded normal human tonsil tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab109531, 1/1000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

       

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D antibody [EP4426] (ab109531)

      ab109531, at 1/100 dilution, staining paraffin embedded Human tonsil tissue by Immunohistochemistry.

      Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

    • Western blot - Anti-CD3D antibody [EP4426] (ab109531)
      Western blot - Anti-CD3D antibody [EP4426] (ab109531)
      All lanes : Anti-CD3D antibody [EP4426] (ab109531) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HuT 78 cell lysate
      Lane 3 : Human thymus tissue lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 19 kDa

    • Anti-CD3D antibody [EP4426] (ab109531)
      Anti-CD3D antibody [EP4426] (ab109531)

    Protocols

    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (6)

    Publishing research using ab109531? Please let us know so that we can cite the reference in this datasheet.

    ab109531 has been referenced in 6 publications.

    • 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
    • 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

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

    Flow Cytometry abreview for Anti-CD3 antibody [EP4426]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Fixation
    Paraformaldehyde
    Sample
    Human Cell (Jurkat (T lymphocytes))
    Specification
    Jurkat (T lymphocytes)
    Permeabilization
    Yes - SAPONIN
    Read More

    Ms. Beatriz Garcillan

    Verified customer

    Submitted Oct 15 2013

    Question

    Antikörper, der humane T Zellen erkennt aber NICHT T Zellen aus der Ratte in IHC-P.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 22 2011

    Answer

    Vielen dank fuer Ihren Anruf. Ich habe nun ein paar Antikoerper fuer Sie herrausgesucht, die nur humane T Zellen erkennen und nicht T Zellen aus der Ratte und fuer IHC-P getestet und garantiert sind. Bitte folgen Sie den jeweiligen Links um zu den Datenblaetern mit mehr Informatinen zugelangen. ab32327 Vielen Dank für Ihren Anruf. Ich habe nun ein paar Antikörper für Sie herausgesucht, die nur humane T Zellen erkennen und nicht T Zellen aus der Ratte und für IHC-P getestet und garantiert sind. Bitte folgen Sie den jeweiligen Links um zu den Datenblättern mit mehr Informationen zugelangen. ab32327 Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-E292-ab32327.html). ab33524 Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-PS1-ab33524.html). ab52959 Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-EP449E-ab52959.html). ab109531 Click here (or use the following: https://www.abcam.com/CD3D-antibody-EP4426-ab109531.html). Ich hoffe, diese Liste hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder zu melden, falls Sie weitere Fragen haben. Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-E292-ab32327.html). ab33524 Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-PS1-ab33524.html). ab52959 Click here (or use the following: https://www.abcam.com/CD3-epsilon-antibody-EP449E-ab52959.html). ab109531 Click here (or use the following: https://www.abcam.com/CD3D-antibody-EP4426-ab109531.html). Ich hoffe, diese Liste hilft Ihnen weiter. Bitte zoegern Sie nicht, sich wioeder zu melden, falls Sie weitere Fragen haben.

    Read More

    Abcam Scientific Support

    Answered on Sep 22 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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