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    cd8-alpha-antibody-epr21769-bsa-and-azide-free-ab230156.pdf

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Immunology Adaptive Immunity T Cells Cytotoxic Cells
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RecombinantRabMAb

Recombinant Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

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Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
  • Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

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 - BSA and Azide free
  • Suitable for: IHC-P, WB, IHC-Fr, Flow Cyt, IP
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 APC FITC Unconjugated

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Overview

  • Product name

    Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free
    See all CD8 alpha primary antibodies
  • Description

    Rabbit monoclonal [EPR21769] to CD8 alpha - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Mouse spleen tissue.
  • General notes

    ab230156 is the carrier-free version of ab217344.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21769
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Cytotoxic Cells
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Cancer
    • Tumor immunology
    • CD markers

Associated products

  • Alternative Versions

    • Anti-CD8 alpha antibody [EPR21769] (ab217344)
    • Alexa Fluor® 488 Anti-CD8 alpha antibody [EPR21769] (ab237364)
    • Alexa Fluor® 647 Anti-CD8 alpha antibody [EPR21769] (ab237365)
    • FITC Anti-CD8 alpha antibody [EPR21769] (ab237367)
    • APC Anti-CD8 alpha antibody [EPR21769] (ab237368)
    • Alexa Fluor® 594 Anti-CD8 alpha antibody [EPR21769] (ab277939)
    • Alexa Fluor® 555 Anti-CD8 alpha antibody [EPR21769] (ab280863)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human CD8 alpha protein (ab114161)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab230156 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
IHC-P
Use at an assay dependent concentration. 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.

WB
Use at an assay dependent concentration. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa).
IHC-Fr
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Notes
IHC-P
Use at an assay dependent concentration. 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.

WB
Use at an assay dependent concentration. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa).
IHC-Fr
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.

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.
  • Target information above from: UniProt accession P01732 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 12525 Mouse
    • SwissProt: P01731 Mouse
    • Unigene: 1858 Mouse
    • 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] - BSA and Azide free (ab230156)
      Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      Clone EPR21769 (ab230156) has been successfully conjugated by Abcam. This image was generated using Anti-CD8 alpha antibody [EPR21769] (Alexa Fluor® 647). Please refer to ab237365 for protocol details.

      IHC image of CD8 alpha staining in a section of formalin-fixed paraffin-embedded normal mouse spleen.

      The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins, performed on a Leica BOND™. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab237365 at 1/100 dilution (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      Clone EPR21769 (ab230156) has been successfully conjugated by Abcam. This image was generated using Anti-CD8 alpha antibody [EPR21769] (Alexa Fluor® 488). Please refer to ab237364 for protocol details.

      IHC image of CD8 alpha staining in a section of formalin-fixed paraffin-embedded normal mouse spleen.

      The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins, performed on a Leica BOND™. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab237364 at 1/100 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

    • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

    • Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

      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.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217344).

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
      Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)

    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

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab230156? Please let us know so that we can cite the reference in this datasheet.

    ab230156 has not yet been referenced specifically in any publications.

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