Invitrogen™ CD25 Monoclonal Antibody (P4A10), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-164-32	100 Tests

Catalog No. 50-164-32 Supplier Invitrogen™ Supplier No. 50025042

Description

Mouse Monoclonal Antibody

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 lymphocytes. This can be used at 5 μL (0.125 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. eFluor™ 660 is a replacement for Alexa Fluor™ 647. eFluor™ 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

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.

Specifications

• Antigen: CD25
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: P4A10
• Concentration: 5 μL/Test
• Conjugate: eFluor 660
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: IL2RA
• Gene Accession No.: O62802
• Gene Alias: ADA; ADA1; Adenosine aminohydrolase; adenosine deaminase; CD25; CD25 antigen; cytokine receptor; IDDM10; IL 2 RA; IL 2 receptor; IL 2R; IL 2R subunit alpha; IL2 RA; IL2 RA antibody; IL2 receptor; il-2 receptor alpha; IL-2 Receptor alpha chain; IL-2 receptor alpha subunit; IL2 receptor subunit alpha; IL-2 receptor subunit alpha; IL2R; IL-2R alpha; IL-2R alpha chain; IL2R subunit alpha; IL-2R subunit alpha; Il2ra; IL-2RA; IL-2-RA; IL2-RA; Il2ra antibody; IL2RAC; IMD41; interleukin 2 receptor; interleukin 2 receptor alpha chain; interleukin 2 receptor subunit alpha; interleukin 2 receptor, alpha; interleukin 2 receptor, alpha chain; interleukin-2 receptor alpha; interleukin-2 receptor alpha chain; interleukin-2 receptor alpha subunit; interleukin-2 receptor beta chain; Interleukin2 receptor subunit alpha; interleukin-2 receptor subunit alpha; Ly-43; p55; p55 chain; sIL 2R; soluble IL 2 receptor; TAC antigen; TCGFR
• Gene Symbols: IL2RA
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 403870
• Target Species: Canine
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

For Research Use Only.