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Anti cd25 pe cy5

Manufactured by Thermo Fisher Scientific
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The Anti-CD25-PE-Cy5 is a fluorescent-labeled antibody that binds to the CD25 antigen, which is expressed on the surface of activated T cells and regulatory T cells. It is designed for use in flow cytometry applications to detect and quantify CD25-positive cell populations.

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Lab products found in correlation

Foxp3 staining buffer kit, by Thermo Fisher Scientific (1 mentions) Live dead fixable blue dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Anti cd4 qdot655, by Thermo Fisher Scientific (1 mentions) Anti foxp3 pe clone pch101, by Thermo Fisher Scientific (1 mentions) Anti cd3 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti ctla 4 apc, by BD (1 mentions) Anti cd39 fitc, by Thermo Fisher Scientific (1 mentions) Anti helios fitc, by BioLegend (1 mentions) Lsrfortessa, by BD (1 mentions) Anti foxp3 apc, by Thermo Fisher Scientific (1 mentions) Anti ctla 4 pe, by Thermo Fisher Scientific (1 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Anti cd4 fluorescein isothiocyanate fitc, by Thermo Fisher Scientific (1 mentions) Anti cd38 pe cy5, by BioLegend (1 mentions) Anti cd19 pe, by Beckman Coulter (1 mentions) Fixation permeabilization buffer solution, by Thermo Fisher Scientific (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions)

3 protocols using anti cd25 pe cy5

1

Comprehensive Treg Immunophenotyping Protocol

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Cryopreserved expanded Tregs and Tconvs were thawed and immunostained with anti-CD3-PE-Cy7, anti-CD4-qdot655 (Invitrogen), anti-CD25-PE-Cy5 (eBiosciences), anti-CD39-FITC (eBioscience), anti-CD45RA-horizon v450 (BD Pharmingen), anti-FOXP3-PE (clone PCH101, eBiosciences), anti-CTLA4-APC (BD Pharmingen), anti-HELIOS-FITC (Biolegend). For intracellular staining, the eBioscience FOXP3 staining buffer kit was used. Dead cells were eliminated using the LIVE/DEAD® Fixable Blue Dead Cell Stain Kit (Invitrogen). Flow-cytometry data were acquired on a LSR Fortessa (BD Biosciences).
Angin M., Klarenbeek P.L., King M., Sharma S.M., Moodley E.S., Rezai A., Piechocka-Trocha A., Toth I., Chan A.T., Goulder P.J., Ndung'u T., Kwon D.S, & Addo M.M. (2014). Regulatory T Cells Expanded from HIV-1-Infected Individuals Maintain Phenotype, TCR Repertoire and Suppressive Capacity. PLoS ONE, 9(2), e86920.
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2

Flow Cytometric Analysis of Regulatory T Cells

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Expanded cells were characterized by staining with anti-CD4 Alexa 700 (BD Pharmigen) and anti-CD25 PE-Cy5 (eBioscience) antibodies. Cells were then permeabilized using the Foxp3 Staining Buffer Set (eBioscience) and stained with anti-Foxp3 APC (eBioscience) and anti-CTLA-4 PE (eBioscience) antibodies. Stained cells were analyzed on an LSRII Flow Cytometer (BD Biosciences) and data compiled on FlowJo software (TreeStar).
Peripheral blood lymphocytes from experimental mice were stained for flow cytometry. After lysis of red blood cells with ACK buffer, cells were permeabilized, stained, and analyzed as described above. Detailed descriptions of analysis techniques are provided in Supplemental Methods.
Smith B.M., Lyle M.J., Chen A.C, & Miao C.H. (2019). Antigen-Specific In vitro Expansion of Factor VIII-Specific Regulatory T Cells Induces Tolerance in Hemophilia A Mice. Journal of thrombosis and haemostasis : JTH, 18(2), 328-340.
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3

Phenotyping of Regulatory T and B Cells

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Approximately 1x10 6 PBMC were deposited into flow cytometry tubes (Falcon, Becton Dickinson, Oxnard, California). To identify Treg cells, surface labeling was performed with anti-CD4-fluorescein isothiocyanate (FITC) (clone: OKT4, eBioscience, San Diego, CA) and anti-CD25-PECy5 (clone: BC96 eBioscience) antibodies. The cells were fixed with 2% paraformaldehyde, permeabilized with the fixation/permeabilization buffer solution (eBioscience, San Diego, CA) according to the manufacturer's recommendations, and subsequently incubated with anti-FOXP3-PE antibody (clone: 259D Biolegend). Finally, the cells were washed and resuspended in phosphate-buffered saline (PBS) and read on an Accuri C6 flow cytometer (BD Biosciences). For the phenotyping of the Breg cells, anti-CD19-PE (Beckman Coulter, Brea, CA, USA), anti-CD24-Alexa-fluor-647 (clone: ML5 BioLegend) and anti-CD38-PECy5 (clone: HIT2 BioLegend) antibodies were added for 30 minutes at room temperature.
Subsequently, the cells were washed with PBS and fixed with 2% paraformaldehyde. The samples were read in a flow cytometer.
Burbano Y.C.B., Paz A.V.C., Caldon C.C.R., Constain J.S.R., Gonzáles G.I.Á., Hernández J.C.K., Marin-Agudelo N., Dueñas-Cuellar R.A, & Castaño V.E.N. (2019). Low Frequency Of Regulatory B-Cells And Increased CD4+ and CD8+ Interferon-γ-producing cells in patients with tropical spastic paraparesis associated with human T-cell lymphotropic virus type. Revista da Sociedade Brasileira de Medicina Tropical, 52.
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