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    cd69-antibody-epr21814-ab233396.pdf

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Immunology Adaptive Immunity T Cells CD
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RecombinantRabMAb

Recombinant Anti-CD69 antibody [EPR21814] (ab233396)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (4) Submit a question References (2)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)
  • Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
  • Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)
  • Immunoprecipitation - Anti-CD69 antibody [EPR21814] (ab233396)
  • Anti-CD69 antibody [EPR21814] (ab233396)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21814] to CD69
  • Suitable for: IP, IHC-P, WB
  • Reacts with: Human

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Carrier Free

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Overview

  • Product name

    Anti-CD69 antibody [EPR21814]
    See all CD69 primary antibodies
  • Description

    Rabbit monoclonal [EPR21814] to CD69
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: PMA (ab120297) treated Daudi whole cell lysate. HL-60, K562, U937 and H9 whole cell lysate. Human tonsil, lymphoma, thymus and colon tissue lysate. IP: PMA (ab120297) treated Daudi whole cell lysate. IHC-P: Human cervix cancer and tonsil tissue.
  • 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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21814
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Immunology
    • Cell Type Markers
    • CD
    • NK Cells
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • NK Cell Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • Alternative Versions

    • Anti-CD69 antibody [EPR21814] - BSA and Azide free (ab234512)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunohistochemistry reagents

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

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

    • Recombinant Human CD69 protein (ab174004)
  • Related Products

    • Phorbol 12-myristate 13-acetate (PMA), PKC activator (ab120297)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab233396 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
IP
1/30.
IHC-P (4)
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 23 kDa.
Notes
IP
1/30.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 23 kDa.

Target

  • Function

    Involved in lymphocyte proliferation and functions as a signal transmitting receptor in lymphocytes, natural killer (NK) cells, and platelets.
  • Tissue specificity

    Expressed on the surface of activated T-cells, B-cells, natural killer cells, neutrophils, eosinophils, epidermal Langerhans cells and platelets.
  • Sequence similarities

    Contains 1 C-type lectin domain.
  • Developmental stage

    Earliest inducible cell surface glycoprotein acquired during lymphoid activation.
  • Post-translational
    modifications

    Constitutive Ser/Thr phosphorylation in both mature thymocytes and activated T-lymphocytes.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession Q07108 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 969 Human
    • Omim: 107273 Human
    • SwissProt: Q07108 Human
    • Unigene: 208854 Human
    • Alternative names

      • Activation inducer molecule (AIM/CD69) antibody
      • Activation inducer molecule antibody
      • AIM antibody
      • BL-AC/P26 antibody
      • BLAC/P26 antibody
      • C-type lectin domain family 2 member C antibody
      • CD69 antibody
      • CD69 antigen (p60, early T-cell activation antigen) antibody
      • CD69 antigen antibody
      • CD69 molecule antibody
      • CD69_HUMAN antibody
      • CLEC2C antibody
      • EA1 antibody
      • Early activation antigen CD69 antibody
      • Early lymphocyte activation antigen antibody
      • Early T cell activation antigen p60 antibody
      • Early T-cell activation antigen p60 antibody
      • GP32/28 antibody
      • Leu23 antibody
      • Leukocyte surface antigen Leu-23 antibody
      • MLR-3 antibody
      • MLR3 antibody
      • VEA antibody
      • Very Early Activation Antigen antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD69 with ab233396 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in lymphoid cells of human tonsil (PMID: 2807372) is observed. Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
      Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
      All lanes : Anti-CD69 antibody [EPR21814] (ab233396) at 1/5000 dilution

      Lane 1 : Un-treated Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate
      Lane 2 : Daudi treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 23 kDa
      Observed band size: 28,32 kDa why is the actual band size different from the predicted?


      Exposure time: 92 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      PMA treatment increases the basal level of p28/32 on Daudi. PMID: 1617156.

    • Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
      Western blot - Anti-CD69 antibody [EPR21814] (ab233396)
      All lanes : Anti-CD69 antibody [EPR21814] (ab233396) at 1/1000 dilution

      Lane 1 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast) whole cell lysate
      Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 3 : U937 (human histiocytic lymphoma monocyte) whole cell lysate
      Lane 4 : Human tonsil
      Lane 5 : Human lymphoma
      Lane 6 : H9 (human lymphoma T lymphocyte) whole cell lysate
      Lane 7 : Human thymus
      Lane 8 : Human colon

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 23 kDa
      Observed band size: 24, 32 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1-5 and 7-8: 3 minutes; Lane 6: 26 seconds.

      We observe an extra band at approximately 15 kDa that we do not yet have an explanation for; it could be non-specific. 

      This blot was developed using a higher sensitive ECL substrate. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)

      Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling CD69 with ab233396 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in infiltrated lymphoid cells of human cervical cancer is observed. Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunoprecipitation - Anti-CD69 antibody [EPR21814] (ab233396)
      Immunoprecipitation - Anti-CD69 antibody [EPR21814] (ab233396)

      CD69 was immunoprecipitated from 0.35 mg Daudi (human Burkitt's lymphoma lymphoblast) treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours whole cell lysate with ab233396 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233396 at 1/5,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.

      Lane 1: Daudi (human Burkitt's lymphoma lymphoblast) treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours whole cell lysate 10 µg (Input).
      Lane 2: ab233396 IP in Daudi treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours whole cell lysate (+).
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233396 in Daudi treated with 50 ng/ml phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours whole cell lysate (-).

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-CD69 antibody [EPR21814] (ab233396)
      Anti-CD69 antibody [EPR21814] (ab233396)

    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

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

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

    References (2)

    Publishing research using ab233396? Please let us know so that we can cite the reference in this datasheet.

    ab233396 has been referenced in 2 publications.

    • Taylor SA  et al. Transcriptional profiling of pediatric cholestatic livers identifies three distinct macrophage populations. PLoS One 16:e0244743 (2021). PubMed: 33411796
    • Li B  et al. Fresh Tissue Multi-omics Profiling Reveals Immune Classification and Suggests Immunotherapy Candidates for Conventional Chondrosarcoma. Clin Cancer Res N/A:N/A (2021). PubMed: 34426437

    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-CD69 antibody [EPR21814]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Eye)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6 & TRIS pH 9
    Permeabilization
    Yes - 0.3% Triton X-100
    Specification
    Eye
    Blocking step
    Donkey Serum 10% + 3% BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Fixative
    Paraformaldehyde
    Read More

    DR. Dylan Windell

    Verified customer

    Submitted May 06 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD69 antibody [EPR21814]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Brain Cortex- Astrocytoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ph9
    Permeabilization
    Yes - 0.1% Triton
    Specification
    Brain Cortex- Astrocytoma
    Blocking step
    Donkey Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    formalin
    Read More

    MRS. Samantha Calderazzo

    Verified customer

    Submitted Sep 24 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD69 antibody [EPR21814]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Leica ER2
    Permeabilization
    No
    Specification
    lymph node
    Fixative
    Formaldehyde
    Read More

    Mr. 勝啓 平田

    Verified customer

    Submitted Apr 05 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD69 antibody [EPR21814]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH6
    Permeabilization
    No
    Specification
    Tonsil
    Blocking step
    Bloxall Endogenous Peroxidase and Alkaline Phosphatase Blocking as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Mr. Joshua Dodge

    Verified customer

    Submitted Feb 25 2019

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