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Invitrogen
CD25 Monoclonal Antibody (PC61.5), NovaFluor™ Blue 585, eBioscience™
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Application
FIGURE: 1 / 2
CD25 Antibody (M060T03B04-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.1 µg of CD25 Monoclonal Antibody, NovaFluor Blue 585 (right). All cells were co-stained with CD4 Monoclonal Antibody, eFluor 450 (Product # 48-0041-82). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
M060T03B04-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG1, lambda
Class
Monoclonal
Type
Antibody
Clone
PC61.5
Conjugate
Excitation/Emission Max
493/586 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3666480
Product Specific Information
Description
The PC61.5 antibody reacts with mouse CD25, the 55 kDa interleukin-2 receptor alpha chain (IL-2R alpha). CD25 is expressed by early progenitors of the 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. Binding of IL-2 to both the high and low affinity classes of IL-2 receptor is inhibited by the PC61.5 antibody. CD25 plays a role in lymphocyte differentiation and activation/proliferation.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This PC61.5 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to (0.1 µg)/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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 493 nm; Emission: 586 nm; Laser: 488 nm (Blue) Laser
Target Information
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.
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Bioinformatics
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-2-RA; IL-2R alpha chain; IL-2R subunit alpha; IL2 RA; IL2 RA antibody; IL2 receptor; IL2 receptor subunit alpha; IL2-RA; IL2R subunit alpha; Il2ra antibody; interleukin-2 receptor alpha; Interleukin-2 receptor subunit alpha; Interleukin2 receptor subunit alpha; p55; p55 chain; sIL 2R; soluble IL 2 receptor
Gene Aliases: CD25; Il2r; Il2ra; Ly-43
UniProt ID: (Mouse) P01590
Entrez Gene ID: (Mouse) 16184
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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