Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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

    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

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)

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