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AB325108

Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81]

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Rabbit Recombinant Monoclonal CD69 antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, Flow Cyt and reacts with Human, Mouse samples.

View Alternative Names

CD69, CLEC2C, Early activation antigen CD69, Activation inducer molecule, BL-AC/P26, C-type lectin domain family 2 member C, EA1, Early T-cell activation antigen p60, GP32/28, Leukocyte surface antigen Leu-23, MLR-3, AIM

4 Images
Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling CD69 with ab325108 at 1/50 (10.0 ug/ml) dilution (Magenta).

Confocal image showing weak cytoplasmic staining in subset of human PBMCs, the expression increased after treatment with ionomycin (500 ng/ml) and PMA (10 ng/ml) for 3 hours.
The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Green). The Nuclear counterstain was DAPI (Blue).

Flow Cytometry - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)
  • Flow Cyt

Supplier Data

Flow Cytometry - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) treated with 500ng/ml ionomycin and 10ng/ml PMA for 3 hours (Lower left and right) / Untreated control (Upper left and right) cells labelling CD69 with ab325108 at 1/500 dilution (0.1ug) / Upper right and Lower right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were stained with anti-CD4 conjugated to Brilliant Violet 421.
Gated on viable lymphoid Cells.

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labelling CD69 with ab325108 at 1/50 (10.0 ug/ml) dilution (Magenta).

Confocal image showing weak cytoplasmic staining in subset of mouse PBMCs, the expression increased after treatment with ionomycin (500 ng/ml) and PMA (10 ng/ml) for 3 hours.
The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Green). The Nuclear counterstain was DAPI (Blue).

Flow Cytometry - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)
  • Flow Cyt

Supplier Data

Flow Cytometry - Alexa Fluor® 647 Anti-CD69 antibody [EPR25398-81] (AB325108)

Flow cytometric analysis of Mouse PBMC (mouse peripheral blood mononuclear cell) treated with 500ng/ml ionomycin and 10ng/ml PMA for 24 hours (Lower left and right) / Untreated control (Upper left and right) cells labelling CD69 with ab325108 at 1/500 dilution (0.1ug) / Upper right and Lower right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were stained with anti-CD3 conjugated to Alexa Fluor® 647.
Gated on viable cells.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR25398-81

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human, Mouse

Applications

ICC/IF, Flow Cyt

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "<p></p>", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "1/500", "FlowCyt-species-notes": "<p></p>" }, "Mouse": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/50", "ICCIF-species-notes": "<p></p>", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "1/500", "FlowCyt-species-notes": "<p></p>" } } }
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Product details

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: 68% PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle|Store in the dark
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

CD69 also known as Activation-Induced C-type Lectin or Early T-cell Activation Antigen is a type II transmembrane protein with a mass of approximately 28-32 kDa. It is expressed mainly on activated leukocytes including T cells B cells and natural killer (NK) cells. Expression occurs rapidly after cell activation making CD69 a robust early marker used in flow cytometry analysis like with CD69 FITC or CD69 PE conjugates to study immune cell activation. The mouse version of CD69 serves as an important model for understanding its function in human biology.
Biological function summary

CD69 functions as a signaling molecule during the immune response influencing both cell proliferation and cytokine production. It is a member of the C-type lectin superfamily and operates as a homodimer on the cell surface. CD69 helps modulate the migration of T cells to inflammatory sites by affecting sphingosine-1-phosphate receptor 1 (S1P1) downregulation directing their tissue retention and primary response roles.

Pathways

The protein plays an important role in the regulation of the NF-kB and JAK-STAT signaling pathways. These are important for immune cell communication and response and CD69 interacts with proteins like the IL-2 receptor complex in these pathways. Additionally its cross-linking can lead to altered expression of cytokines and chemokines impacting overall immune function.

Altered CD69 expression links to conditions like autoimmune diseases and chronic inflammation. Its involvement in rheumatoid arthritis shows how CD69 may regulate abnormal immune responses. Furthermore the modulation of CD69 expression can be associated with cancer progression highlighting connections to proteins like FN50 and FN61 which participate in tumor immunity and metastasis control.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Transmembrane protein expressed mainly on T-cells resident in mucosa that plays an essential role in immune cell homeostasis. Rapidly expressed on the surface of platelets, T-lymphocytes and NK cells upon activation by various stimuli, such as antigen recognition or cytokine signaling, stimulates different signaling pathways in different cell types (PubMed : 24752896, PubMed : 26296369, PubMed : 35930205). Negatively regulates Th17 cell differentiation through its carbohydrate dependent interaction with galectin-1/LGALS1 present on immature dendritic cells (PubMed : 24752896). Association of CD69 cytoplasmic tail with the JAK3/STAT5 signaling pathway regulates the transcription of RORgamma/RORC and, consequently, differentiation toward the Th17 lineage (By similarity). Acts also via the S100A8/S100A9 complex present on peripheral blood mononuclear cells to promote the conversion of naive CD4 T-cells into regulatory T-cells (PubMed : 26296369). Acts as an oxidized low-density lipoprotein (oxLDL) receptor in CD4 T-lymphocytes and negatively regulates the inflammatory response by inducing the expression of PDCD1 through the activation of NFAT (PubMed : 35930205). Participates in adipose tissue-derived mesenchymal stem cells (ASCs)-mediated protection against P. aeruginosa infection. Mechanistically, specifically recognizes P. aeruginosa to promote ERK1 activation, followed by granulocyte-macrophage colony-stimulating factor (GM-CSF) and other inflammatory cytokines secretion (PubMed : 34841721). In eosinophils, induces IL-10 production through the ERK1/2 pathway (By similarity). Negatively regulates the chemotactic responses of effector lymphocytes and dendritic cells (DCs) to sphingosine 1 phosphate/S1P by acting as a S1PR1 receptor agonist and facilitating the internalization and degradation of the receptor (PubMed : 37039481).
See full target information CD69
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com