Rabbit anti-Human/Monkey CD69 Monoclonal Antibody for Flow Cytometry - Q07108

Core Product Specifications and Parameters

Biological Background: CD69 Function and Localization

• CD69 (Cluster of Differentiation 69) is a transmembrane glycoprotein and C-type lectin domain family 2 member C (CLEC2C), also known as activation inducer molecule (AIM) or early T-cell activation antigen p60.
• It is rapidly expressed on the surface of platelets, T-lymphocytes, B-cells, natural killer (NK) cells, neutrophils, eosinophils, epidermal Langerhans cells, and monocytes upon activation by antigen recognition or cytokine signaling.
• CD69 plays an essential role in immune cell homeostasis and negatively regulates Th17 cell differentiation through carbohydrate-dependent interaction with galectin-1/LGALS1 on immature dendritic cells. Related references: PMID:24752896
• It promotes the conversion of naive CD4+ T-cells into regulatory T-cells via the S100A8/S100A9 complex on peripheral blood mononuclear cells. Related references: PMID:26296369
• CD69 acts as an oxidized low-density lipoprotein (oxLDL) receptor in CD4 T-lymphocytes and negatively regulates the inflammatory response by inducing PDCD1 expression through NFAT activation. Related references: PMID:35930205
• In adipose tissue-derived mesenchymal stem cell-mediated protection against P. aeruginosa infection, CD69 specifically recognizes the pathogen to promote ERK1 activation and cytokine secretion. Related references: PMID:34841721
• It negatively regulates chemotactic responses to sphingosine 1-phosphate (S1P) by acting as a S1PR1 receptor agonist and facilitating receptor internalization and degradation. Related references: PMID:37039481
• Subcellularly, CD69 is localized to the cell membrane and contains a signal-anchor, a transmembrane helix, and a C-type lectin domain; it undergoes phosphorylation and N-linked glycosylation.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 78-199 of human CD69, covering the C-type lectin domain; ensure epitope accessibility under native conditions for flow cytometry.
• For flow cytometry, titrate the antibody to an optimal concentration using a relevant positive control (e.g., activated human PBMCs) and include fluorescence-minus-one (FMO) controls.
• Consider fixation and permeabilization only if intracellular staining is required, as CD69 is predominantly surface-expressed; validate in your specific assay system.
• Note that CD69 is transiently expressed; synchronization of cell activation timing may improve staining consistency.
• Cross-reactivity with cynomolgus monkey has been confirmed; for other species, test empirically.

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