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  1. Link

    cd8-alpha-antibody-sp16-ab101500.pdf

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Immunology Adaptive Immunity T Cells Cytotoxic Cells
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RecombinantRabMAb

Recombinant Anti-CD8 alpha antibody [SP16] (ab101500)

  • Datasheet
  • SDS
Reviews (3)Q&A (1)References (24)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)
  • Flow Cytometry - Anti-CD8 alpha antibody [SP16] (ab101500)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP16] to CD8 alpha
  • Suitable for: IHC-P, Flow Cyt
  • Reacts with: Human

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Overview

  • Product name

    Anti-CD8 alpha antibody [SP16]
    See all CD8 alpha primary antibodies
  • Description

    Rabbit monoclonal [SP16] to CD8 alpha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD8 alpha aa 200 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P01732

  • Epitope

    C-terminus
  • Positive control

    • IHC-P: Human tonsil and lymph node tissue. Flow Cyt: Human peripheral blood lymphocytes.
  • 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.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.2
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    SP16
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Cytotoxic Cells
    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Cancer
    • Tumor immunology
    • CD markers
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab101500 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 (3)
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

Flow Cyt
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Notes
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

Flow Cyt
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

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: 925 Human
    • Omim: 186910 Human
    • SwissProt: P01732 Human
    • Unigene: 85258 Human
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)

      Immunohistochemical analysis of paraffin-embedded Human tonsil labeling CD8 with ab101500 at 1/100 (21 µg/ml). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab101500 for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. DAB was used as the chromogen. Counterstained with Hematoxylin and mounted with DPX.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [SP16] (ab101500)Image from Graff JN et al.,Oncotarget 7(33), 52810 - 52817. Fig 2,; doi: 10.18632/oncotarget.10547. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/3.0/.

      This image was generated using a previous batch manufactured using hybridoma production method.

      IHC using multi-spectral imaging on human lymph node (A-C) obtained from men with mCRPC. A) H+E staining and B) single-color images (plus nuclear stain; DAPI) of CD3 (ab16669), CD8 alpha (ab101500), CD163, PD-L1, cytokeratin (CK), DAPI and C) merged.  H+E stating at 20X magnification; multi-spectral images 200X magnification.

    • Flow Cytometry - Anti-CD8 alpha antibody [SP16] (ab101500)
      Flow Cytometry - Anti-CD8 alpha antibody [SP16] (ab101500)

      This image was generated using a previous batch manufactured using hybridoma production method.

      Human peripheral blood lymphocytes staining CD8 alpha with ab101500 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab101500, 1/1000 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (24)

    Publishing research using ab101500? Please let us know so that we can cite the reference in this datasheet.

    ab101500 has been referenced in 24 publications.

    • Moreira D  et al. Myeloid cell-targeted STAT3 inhibition sensitizes head and neck cancers to radiotherapy and T cell-mediated immunity. J Clin Invest 131:N/A (2021). PubMed: 33232304
    • Cui C  et al. Propofol maintains Th17/Treg cell balance and reduces inflammation in rats with traumatic brain injury via the miR-145-3p/NFATc2/NF-?B axis. Int J Mol Med 48:N/A (2021). PubMed: 34036377
    • Wang G  et al. The pan-cancer landscape of crosstalk between epithelial-mesenchymal transition and immune evasion relevant to prognosis and immunotherapy response. NPJ Precis Oncol 5:56 (2021). PubMed: 34158591
    • Roelants C  et al. Ex-Vivo Treatment of Tumor Tissue Slices as a Predictive Preclinical Method to Evaluate Targeted Therapies for Patients with Renal Carcinoma. Cancers (Basel) 12:N/A (2020). PubMed: 31963500
    • Ji D  et al. Combination of radiotherapy and suppression of Tregs enhances abscopal antitumor effect and inhibits metastasis in rectal cancer. J Immunother Cancer 8:N/A (2020). PubMed: 33106387
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 antibody [SP16]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (NSCLC)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH6 (3 min 1000W, 30 min 100W)
    Permeabilization
    No
    Specification
    NSCLC
    Blocking step
    Akoya Biosciences 7 color OPAL Kit Blocking solution as blocking agent for 10 minute(s) · Concentration: 100%
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 04 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 antibody [SP16]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Cynomolgus monkey Tissue sections (Thymus)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA pH 9.0
    Permeabilization
    No
    Specification
    Thymus
    Blocking step
    Casein as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 24 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 antibody [SP16]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    2.5% Normal Horse Serum - VectorLabs ImmPRESS™ Excel Amplified HRP Polymer Staining Kit (Anti-Mouse Ig) as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: RT°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA pH 9.0 Bond Epitope Retrieval Solution 2 (ER2, Leica)
    Sample
    Rhesus monkey Tissue sections (Spleen)
    Specification
    Spleen
    Permeabilization
    No
    Fixative
    10% Neutral Buffered Formalin
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 26 2015

    Question

    Hi, Do you know if the above CD8 cross-reacts on monkey FFPE tissues? Would you be able to provide the concentration for the 100 uL size?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 04 2014

    Answer

    The CD8 antibody ab101500 is produced from tissue culture supernatants and the IgG concentration is not determined. We do not have any experimental data regarding the cross-reactivity of ab101500 with monkey samples. The immunogen for the antibody has 84% homology with the rhesus sequence below, so reactivity with that species is possible.

    MAPPVTALLLPLVLLLHAARPNQFRVSPLGRTWNLGETVELKCQVLLSNPTSGCSWLFQP
    RGTAARPTFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLRDFRQENEGYYFCSALSN
    SIMYFSHFVPVFLPAKPTTTPAPRSPTPAPTTASQPLSLRPEACRPAAGGSVNTRGLDFA
    CDIYIWAPLAGACGVLLLSLVITLYCNHRNRRRVCKCPRPVVKSGGKPSLSDRYV

    http://www.uniprot.org/uniprot/F7DXL7.fasta

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Apr 04 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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