Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) treated with 10μg/ml PHA for 24h labelling CD25/IL-2Rα antibody at 1/200 (1 μg) dilution (Right) compared with a Mouse monoclonal IgG isotype control (Left). Goat Anti - Mouse IgG Alexa Fluor® 647 was used as the secondary antibody.
Then cells were stained with CD4 - Phycoerythrin separately.
CD25/IL-2Rα Mouse mAb (S-573-1)
CD25/IL-2Rα Mouse mAb (S-573-1)
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Product Details
Product Details
Product Specification
| Host | Mouse |
| Antigen | CD25/IL-2Rα |
| Synonyms | Interleukin-2 receptor subunit alpha, IL-2 receptor subunit alpha, IL-2-RA, IL-2R subunit alpha, IL2-RA, TAC antigen, p55 |
| Immunogen | Recombinant Protein |
| Location | Membrane |
| Accession | P01589 |
| Clone Number | S-573-1 |
| Antibody Type | Mouse mAb |
| Isotype | IgG1,k |
| Application | ICC, FCM |
| Reactivity | Hu |
| Purification | Protein G |
| Concentration | 2 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 | |
| FCM | 1:2000 |
Background
CD25 is expressed broadly among leukocytes. The highest surface expression of this protein is on regulatory T cells (Tregs), where CD25 is expressed constitutively, especially on a subset classified as naturally occurring Tregs. It can also be found on activated B cells, NK (natural killer) cells, thymocytes, and some myeloid lineage cells (e.g. macrophages, dendritic cells). L2RA has been used as a marker to identify CD4+FoxP3+ regulatory T cells in mice, and it has been found that a large proportion of resting memory T cells constitutively express IL2RA in humans. Levels of CD25 soluble form, called sIL-2Rα, has been connected to pathogenesis of autoimmune diseases and cancer. Since sCD25 is produced during immune activation, it is used as one of biomarkers to track disease progression and to indicate outcome for clinical disorders. Especially, it is a hallmark for hyper-activated immune system and cytokine storm, which may lead to multiple organ system failure.
Picture
Picture
FC
Immunocytochemistry
ICC shows positive staining in myc-his-IL-2R alpha/CD25 transfected 293T cells. Anti-CD25/IL-2Rα antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Mouse 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).
Negative control: ICC shows negative staining in vector-transfected 293T cells. Anti-CD25/IL-2Rα antibody was used at 1/500 dilution and incubated overnight at 4°C. Goat polyclonal Antibody to Mouse 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).
