Recombinant
RabMAb

Recombinant Anti-CD69 antibody [EPR21814] (ab233396)

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 within Human CD69 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q07108

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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 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.
  • Information by UniProt
  • Database links

  • 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

  • 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).

  • 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.

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

    Lane 1 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast) whole cell lysate at 20 µg
    Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
    Lane 3 : U937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg
    Lane 4 : H9 (human lymphoma T lymphocyte) whole cell lysate at 10 µg
    Lane 5 : Human tonsil at 20 µg
    Lane 6 : Human lymphoma at 20 µg
    Lane 7 : Human thymus at 20 µg
    Lane 8 : Human colon at 20 µg

    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.

  • 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).

  • 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.

References

ab233396 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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

Mr. Joshua Dodge

Verified customer

Submitted Feb 25 2019

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

Sign up