Description: The BC96 monoclonal antibody reacts with human CD25, the 55 kDa interleukin-2 receptor alpha chain (IL-2R alpha). CD25 is expressed by early progenitors of T and B lineage as well as by activated mature T and B lymphocytes. By itself, CD25 binds IL-2 only with low affinity. However, CD25 associates with CD122 (IL-2 receptor beta chain) and CD132 (common gamma chain) to form the high affinity IL-2 receptor. CD25 plays a role in lymphocyte differentiating and activation/proliferation.
Applications Reported: This BC96 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This BC96 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may 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 780 can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
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: 780 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: CD25; 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; interleukin-2 receptor alpha; Interleukin-2 receptor subunit alpha; Interleukin2 receptor subunit alpha; p55; sIL 2R; soluble IL 2 receptor; TAC antigen
Gene Aliases: CD25; IDDM10; IL2R; IL2RA; IMD41; p55; TCGFR
UniProt ID: (Human) P01589
Entrez Gene ID: (Human) 3559