Recombinant
RabMAb

Recombinant Anti-CD8 alpha antibody [EPR21769] (ab217344)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21769] to CD8 alpha
  • Suitable for: WB, IHC-Fr, IHC-P, Flow Cyt, IP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-CD8 alpha antibody [EPR21769]
    See all CD8 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR21769] to CD8 alpha

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Fr, IHC-P, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment within Mouse CD8 alpha aa 1-200. The exact sequence is proprietary.
    Database link: P01731

  • Positive control

    • WB: Mouse thymus, spleen and lymph node lysates. IHC-P: Mouse spleen and colon tissues. IHC-Fr: Mouse spleen and thymus tissues. Flow Cyt: Mouse primary splenocytes. IP: Mouse thymus 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.

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 long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR21769

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab217344 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. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa).
IHC-Fr 1/500.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Background staining was observed in mouse testis samples.
Flow Cyt 1/500.
IP 1/30.

Target

  • Function

    Identifies cytotoxic/suppressor T-cells that interact with MHC class I bearing targets. CD8 is thought to play a role in the process of T-cell mediated killing. CD8 alpha chains binds to class I MHC molecules alpha-3 domains.

  • Involvement in disease

    Defects in CD8A are a cause of familial CD8 deficiency (CD8 deficiency) [MIM:608957]. Familial CD8 deficiency is a novel autosomal recessive immunologic defect characterized by absence of CD8+ cells, leading to recurrent bacterial infections.

  • Sequence similarities

    Contains 1 Ig-like V-type (immunoglobulin-like) domain.

  • Post-translational
    modifications

    All of the five most carboxyl-terminal cysteines form inter-chain disulfide bonds in dimers and higher multimers, while the four N-terminal cysteines do not.

  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • alpha polypeptide (p32) antibody
      • CD_antigen=CD8a antibody
      • CD8 antibody
      • CD8 antigen alpha polypeptide antibody
      • CD8 antigen alpha polypeptide (p32) antibody
      • CD8 antigen, alpha polypeptide (p32) antibody
      • CD8a antibody
      • CD8A antigen antibody
      • CD8A molecule antibody
      • CD8A_HUMAN antibody
      • Leu2 antibody
      • Leu2 T lymphocyte antigen antibody
      • Ly 2 antibody
      • Ly 35 antibody
      • Ly B antibody
      • Ly2 antibody
      • Ly3 antibody
      • Ly35 antibody
      • LyB antibody
      • Lyt 2.1 lymphocyte differentiation antigen (AA at 100) antibody
      • LYT3 antibody
      • MAL antibody
      • OKT8 T cell antigen antibody
      • OTTHUMP00000160760 antibody
      • OTTHUMP00000160764 antibody
      • OTTHUMP00000203528 antibody
      • OTTHUMP00000203721 antibody
      • p32 antibody
      • T cell antigen Leu2 antibody
      • T cell co receptor antibody
      • T lymphocyte differentiation antigen T8/Leu 2 antibody
      • T-cell surface glycoprotein CD8 alpha chain antibody
      • T-cell surface glycoprotein Lyt 2 antibody
      • T-lymphocyte differentiation antigen T8/Leu-2 antibody
      • T8 T cell antigen antibody
      • T8/Leu-2 T-lymphocyte differentiation antigen antibody
      see all

    Images

    • Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      Flow cytometric analysis of mouse primary splenocytes labeling CD8 alpha with ab217344 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

      Cells were surface stained with CD4-Alexa Fluor® 647, then stained with rabbit IgG (Left) / ab217344 (Right) separately. CD4 and CD8 alpha are mutually exclusive expressed in mouse spleen. Gated on total viable cells.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD8 alpha with ab217344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the white pulp of mouse spleen (PMID: 23482450; PMID: 25826597). Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse spleen (PMID: 25616911).

      The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      CD8 alpha was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab217344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217344 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: Mouse thymus lysate 10 µg (Input). 

      Lane 2: ab217344 IP in mouse thymus lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214344 in mouse thymus lysate.

      Exposure time: 5 seconds.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      The two bands are different isoforms that are consistent with the literature (PMID 3085089).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD8 alpha with ab217344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on stromal cells of mouse colon. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

       

    • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)

      Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse thymus tissue section (PMID: 25616911).

      The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Western blot - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      Western blot - Anti-CD8 alpha antibody [EPR21769] (ab217344)
      All lanes : Anti-CD8 alpha antibody [EPR21769] (ab217344) at 1/1000 dilution

      Lane 1 : Mouse thymus lysate at 20 µg
      Lane 2 : Mouse spleen lysate at 20 µg
      Lane 3 : Mouse lymph node lysate at 10 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 27 kDa
      Observed band size: 35 kDa why is the actual band size different from the predicted?



      Exposure time : Lane 1: 3 sconds; Lane 2: 40 seconds; Lane 3: 15 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      The two bands are different isoforms that are consistent with the literature (PMID 3085089).

      The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References

    Publishing research using ab217344? Please let us know so that we can cite the reference in this datasheet.

    ab217344 has been referenced in 5 publications.

    • Amirgholami N  et al. A. crassicauda, M. eupeus and H. lepturus scorpion venoms initiate a strong in vivo anticancer immune response in CT26-tumor mice model. Toxicon 180:31-38 (2020). PubMed: 32275983
    • Chen S  et al. Eluted 25-hydroxyvitamin D3 from radially aligned nanofiber scaffolds enhances cathelicidin production while reducing inflammatory response in human immune system-engrafted mice. Acta Biomater 97:187-199 (2019). PubMed: 31386930
    • Yu J  et al. Genetic Aberrations in the CDK4 Pathway Are Associated with Innate Resistance to PD-1 Blockade in Chinese Patients with Non-Cutaneous Melanoma. Clin Cancer Res 25:6511-6523 (2019). PubMed: 31375512
    • Yu S  et al. A novel asymmetrical anti-HER2/CD3 bispecific antibody exhibits potent cytotoxicity for HER2-positive tumor cells. J Exp Clin Cancer Res 38:355 (2019). PubMed: 31412896
    • Ben Khalaf N  et al. The mouse intron-nested gene, Israa, is expressed in the lymphoid organs and involved in T-cell activation and signaling. Mol Immunol 111:209-219 (2019). PubMed: 31096062

    Customer reviews and Q&As

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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 alpha antibody [EPR21769]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: FLEX Taget Retrieval Solution High pH(Dako k8004) buffer
    Permeabilization
    No
    Specification
    tumor
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 04 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 alpha antibody [EPR21769]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (B16F10(melanoma) syngeneic mouse tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA
    Permeabilization
    Yes - 3% H2O2
    Specification
    B16F10(melanoma) syngeneic mouse tumor
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 29 2019

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