Rabbit anti-Mouse CD69 mAb for Flow Cytometry - P37217

Core Product Specifications and Parameters

Biological Background: CD69 Function and Localization

• Early activation antigen CD69 is a transmembrane C-type lectin receptor that is rapidly upregulated on the surface of platelets, T-lymphocytes, and NK cells upon activation, playing a central role in immune cell homeostasis and signaling.
• Negatively regulates Th17 cell differentiation through carbohydrate-dependent interaction with galectin-1/LGALS1 on immature dendritic cells, and the cytoplasmic tail association with JAK3/STAT5 signaling controls RORγt transcription. Related references: PMID:20696842
• Acts as a negative regulator of lymphocyte and dendritic cell chemotaxis by acting as a sphingosine 1-phosphate receptor 1 (S1PR1) agonist, promoting S1PR1 internalization and degradation. Related references: PMID:16525420
• In eosinophils, CD69 engagement induces IL-10 production through ERK1/2 pathway activation, contributing to immunoregulatory functions. Related references: PMID:38540778
• Also functions as an oxidized low-density lipoprotein (oxLDL) receptor in CD4 T-lymphocytes, negatively regulating inflammatory responses via NFAT-dependent PDCD1 induction, and promotes regulatory T-cell conversion through S100A8/S100A9 complex interaction.
• Localizes to the cell membrane with a single-pass transmembrane topology and a signal-anchor sequence; the extracellular domain includes a carbohydrate recognition domain (CRD) (UniProt: P37217).
• Tissue specificity: detected on the surface of activated T-cells, B-cells, natural killer cells, neutrophils, platelets, and eosinophils.
• Post-translational modifications include disulfide bond formation, N-linked glycosylation, and phosphorylation, as indicated by keywords: "Disulfide bond", "Glycoprotein", "Phosphoprotein".

Experimental Guidance and Technical Tips

• The immunogen used (aa 64–199 of mouse CD69) covers most of the extracellular carbohydrate recognition domain (CRD), which is exposed on the cell surface; this design likely ensures efficient recognition of the native protein in flow cytometry.
• For flow cytometry applications, consider titrating the antibody to determine the optimal staining concentration for your specific cell type and instrument settings.
• Include appropriate controls such as isotype-matched negative controls, fluorescence-minus-one (FMO) controls, and live/dead discrimination to accurately gate on CD69-positive populations.
• Validate specificity by staining cells from CD69 knockout mice or by using a blocking peptide/protein if available, to confirm signal is antigen-dependent.
• Be aware that CD69 is a rapid activation marker; time of stimulation and fixation/permeabilization conditions may need optimization depending on the biological context.

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