ICC analysis of Mouse PBMC cells treated with ionomycin (500 ng/ml,24h) and PMA (10ng/ml ,24h) (top panel) and untreated Mouse PBMC cells (below panel). Anti- CD69 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | CD69 |
| Synonyms | Early activation antigen CD69; Activation inducer molecule (AIM); BL-AC/P26; C-type lectin domain family 2 member C; EA1; Early T-cell activation antigen p60; CLEC2C |
| Location | Cell membrane |
| Accession | Q07108 |
| Clone Number | S-3362 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | ICC |
| Reactivity | Hu, Ms |
| Positive Sample | ionomycin and PMA treated mouse PBMC cells |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| ICC | 1:500 | Ms |
| FCM | 5μl per million cells in 100μl volume |
Background
CD69 is a type II transmembrane glycoprotein and a member of the C-type lectin superfamily. It is a classical early marker of lymphocyte activation, appearing rapidly on the cell surface after stimulation. Structurally, CD69 is a disulfide-linked homodimer protein with two differentially glycosylated subunits (28–32 kDa), each consisting of an extracellular C-type lectin domain connected to a single-spanning transmembrane region followed by a short cytoplasmic tail. It is broadly expressed on the surfaces of most hematopoietic lineages, including activated T and B lymphocytes, natural killer (NK) cells, murine macrophages, neutrophils, and eosinophils, and is constitutively expressed on human monocytes, platelets, epidermal Langerhans cells, and bone-marrow myeloid precursors. CD69 plays various regulatory roles in immune responses, such as promoting tissue residency, regulating Th17/Treg cell differentiation, and contributing to the exhaustion of resident memory T cells, especially in the tumor microenvironment. Its ligands include galectin-1, calprotectin (S100A8/S100A9 complex), myosin light chains 9, 12a, and 12b (Myl9/12), and oxidized low-density lipoprotein (ox-LDL).
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Immunocytochemistry
