Recombinant Anti-CD69 antibody [EPR21814] (ab233396)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-308560-anti-cd69-antibody-epr21814-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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
Related conjugates and formulations
Overview
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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.
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Positive control
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Recombinant Protein
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Related Products
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.
|
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| 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. |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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WB
1/1000. Predicted molecular weight: 23 kDa. |
Target
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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
modificationsConstitutive Ser/Thr phosphorylation in both mature thymocytes and activated T-lymphocytes. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 969 Human
- Omim: 107273 Human
- SwissProt: Q07108 Human
- Unigene: 208854 Human
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Alternative names
- Activation inducer molecule (AIM/CD69) antibody
- Activation inducer molecule antibody
- AIM antibody
see all
Images
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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)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-308558-anti-cd69-antibody-epr21814-western-blot.jpg)
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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
PMA treatment increases the basal level of p28/32 on Daudi. PMID: 1617156.
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![Western blot - Anti-CD69 antibody [EPR21814] (ab233396)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-308559-anti-cd69-antibody-epr21814-western-blot.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD69 antibody [EPR21814] (ab233396)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-308561-anti-cd69-antibody-epr21814-immunohistochemistry.jpg)
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)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-308562-anti-cd69-antibody-epr21814-immunoprecipitation.jpg)
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)](https://www.abcam.com/ps/products/233/ab233396/Images/ab233396-6-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
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