Description: This P4A10 monoclonal antibody reacts with canine CD25, the 55-kDa interleukin-2 receptor alpha (IL-2Ralpha) chain. As in humans and mice, CD25 in dogs is expressed by early T and B lineage progenitors as well as by activated mature T and B lymphocytes. CD25 associates with the IL-2R beta (CD122) and gamma (CD132) chains to form the high-affinity IL-2 receptor. On its own, CD25 binds IL-2 with low affinity. Along with CD4 and Foxp3, CD25 is a major marker used to define regulatory T cells.
The P4A10 antibody has been reported to recognize the same epitope as the anti-human CD25 monoclonal antibody ACT-1, but with greater affinity.
Applications Reported: This P4A10 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This P4A10 antibody has been pre-titrated and tested by flow cytometric analysis of normal canine peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Super Bright 436 can be excited with the violet laser line (405 nm) and emits at 436 nm. We recommend using a 450/50 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 436 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CD25 (IL2 receptor alpha chain/IL2RA) is a cytokine that plays a role in the proliferation of T and B lymphocytes. The receptor of this cytokine (IL2RA) is a heterotrimeric protein complex with a gamma chain also shared by interleukin 4 (IL4) and interleukin 7 (IL7). IL2RA, IL2R beta chain (IL2RB), and the IL2R gamma chain (IL2RG), constitute the high-affinity IL2 receptor. Homodimeric IL2RA chains result in low-affinity receptor, while homodimeric IL2RB chains produce a medium-affinity receptor. The expression of IL2 in mature thymocytes is monoallelic, which represents an unusual regulatory mode for controlling the precise expression of a single gene. IL2 is primarily produced by mature T cells. IL2 plays an important role as a growth factor, differentiation factor, and regulator of cell death. IL-2 stimulates the proliferation of B cells, augments natural killer cell activity, and inhibits granulocyte macrophage colony formation. The targeted disruption of a similar gene in mice leads to ulcerative colitis-like disease, which suggests a role in the immune response to antigenic stimuli. Mutations in this gene are associated with interleukin 2 receptor alpha deficiency.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: IL 2 RA; IL 2 receptor; IL 2R; IL 2R subunit alpha; il-2 receptor alpha; IL-2 receptor subunit alpha; IL-2R subunit alpha; IL2 RA; IL2 RA antibody; IL2 receptor; IL2 receptor subunit alpha; IL2R subunit alpha; Il2ra antibody; interleukin 2 receptor alpha chain; interleukin-2 receptor alpha; Interleukin2 receptor subunit alpha; sIL 2R; soluble IL 2 receptor; TAC antigen
Gene Aliases: CD25; IL-2RA
Entrez Gene ID: (Dog) 403870
Molecular Function: transmembrane signal receptor