RY775 Mouse Anti-Human CD25 (IL-2 Receptor α)

BD Horizon™ RY775 Mouse Anti-Human CD25 (IL-2 Receptor α)

Name: RY775 Mouse Anti-Human CD25 (IL-2 Receptor α)
Brand: RY775 Mouse Anti-Human CD25 (IL-2 Receptor α)
SKU: 571663

Clone M-A251 (RUO)

Flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated and PHA-stimulated Human peripheral blood lymphocytes Left and Middle Plots - Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424) and with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (Cat. No. 571408; Left Plot) or BD Horizon™ RY775 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 571663/571741; Middle Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202).The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Right Plot - Human peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA). The cells were stained with either BD Horizon™ RY775 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ RY775 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (solid line histogram). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD25 (IL-2 Receptor α) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphoblasts. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

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Product Details

Brand: BD Horizon™

Alternative Name: IL-2R; IL2RA; IL-2Rα; TCGFR; TAC antigen; p55

Reactivity: Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Phytohemagglutinin stimulated human lymphocytes

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Workshop Number: IV A053

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
An isotype control should be used at the same concentration as the antibody of interest.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

RY775 Mouse IgG1, κ Isotype Control RUO

Size 0.1 mg Cat No. 571408

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

571663 Rev. 1

Antibody Details

M-A251

The M-A251 monoclonal antibody specifically binds to the 55 kDa type I transmembrane glycoprotein known as low-affinity interleukin-2 receptor alpha chain subunit (IL-2Rα). CD25 is expressed on regulatory T cells, activated lymphocytes (T and B), and monocytes. It associates with the IL-2Rβ/CD122 and IL-2Rγ/CD132 receptor chains to form the high-affinity IL-2R complex. CD25 expression on T and B lymphocytes is upregulated by antigenic or mitogenic stimulation. Soluble CD25/IL-2Rα is produced as a consequence of lymphocyte stimulation and is found in biological fluids following inflammatory responses.

571663 Rev. 1

Format Details

RY775

The BD Horizon RealYellow™ 775 (RY775) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY775 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter).

Format: RY775

Excitation Source: Yellow-Green 561 nm

Excitation Max: 557 nm

Emission Max: 775 nm

571663 Rev.1

Citations & References

View product citations for antibody "571663" on CiteAb

Development References (8)

Herodin F, Thullier P, Garin D, Drouet M. Nonhuman primates are relevant models for research in hematology, immunology and virology. Eur Cytokine Netw. 2005; 16(2):104-116. (Clone-specific: Flow cytometry). View Reference
Janszen M, Buck D, Maino VC. Functional and molecular properties of CD25 monoclonal antibodies. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:403-406.
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
Nakamura K, Kitani A, Fuss I, et al. TGF-β1 plays an important role in the mechanism of CD4+CD25+ regulatory T cell activity in both humans and mice. J Immunol. 2004; 172(2):834-842. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
Ravoet AM, Latinne D, Couvreur B, et al. CD25 mab: Epitopes recognized, effect on lymphocyte activation, mediation of ADCC. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:408-411.
Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
Schwarting R, Stein H. Cluster report: CD25. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:399-403.

View All (8)

571663 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.