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Anti cd25 pe cy7

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Anti-CD25-PE/Cy7 is a monoclonal antibody conjugated with the fluorescent dye PE/Cy7. It is designed to detect and quantify cells expressing the CD25 antigen.

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Lab products found in correlation

Anti cd4 apc cy7, by BioLegend (4 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (3 mentions) Anti cd69 bv510, by BioLegend (2 mentions) Anti foxp3 apc, by Miltenyi Biotec (2 mentions) Anti cd3 af488, by BioLegend (2 mentions) Anti hla dr percp cy5, by BioLegend (2 mentions) Anti cd39 apc, by BioLegend (2 mentions) Guava easycyte 8ht, by Tree Star (2 mentions) Anti foxp3 pe, by BioLegend (2 mentions) Anti cd38 pecy7, by BioLegend (2 mentions) Cytofix cytoperm, by BD (2 mentions) Anti pd 1 pe cy7, by BioLegend (2 mentions) Anti cd19 apc cy7, by BioLegend (2 mentions) Anti cd127 percp cy5, by BioLegend (2 mentions) Anti foxp3 alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Anti cd3 efluor 605nc, by Thermo Fisher Scientific (2 mentions) Anti cd4 fitc, by BioLegend (2 mentions) Anti f4 80 apc, by Thermo Fisher Scientific (2 mentions) Anti cd45 fitc, by Thermo Fisher Scientific (2 mentions) Foxp3 fix perm buffer set, by BioLegend (2 mentions)

Close spelling variants of this product

CD25-PE (55 citations) Anti-CD25-PE (36 citations) CD25 PE-Cy7 (26 citations) PE-Cy7-conjugated anti-CD25 (4 citations) Anti-human CD25 PE-Cy7 (3 citations) PE/Cy7-anti-mouse CD25 (2 citations) PE/Cy7 anti-mouse CD25 antibody (2 citations) PE-conjugated anti-CTLA-4 (UC10-4F10-11); PE-Cy7–conjugated anti-CD25 (PC81); biotin-conjugated anti-CD8b (53-5.8), (2 citations)

15 protocols using anti cd25 pe cy7

1

Multiparametric Flow Cytometry Panel

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The following antibodies were used for flow cytometric stainings anti-TACI PE (clone 1A1), anti-CD19 APC-Cy7, anti-CD27 PerCP-Cy5.5 or APC, anti-CD10 PE-Cy7, anti-IgM FITC, anti-CD21 APC, anti-CD69 PE-Cy7, anti-CD86 PE, anti-CD4 APC-Cy7, anti-CD25 PECy7, anti-CD127 PerCP-Cy5.5, anti-CD45RO Pacific Blue, anti-CXCR5 PerCP-Cy5.5, anti-PD-1 PE-Cy7, anti-CD25 PE, anti-CD25 PE-Cy7 (all from BioLegend, San Diego, Calif), anti-CD3 eFluor 605NC, anti-CD21 BD Horizon V450 (Becton Dickinson) and goat polyclonal anti-TACI biotin (R&D Systems). Intracellular staining with anti-Foxp3 Alexa Fluor 488 and anti-BCL6 PE (eBioscience, San Diego, Calif) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), in accordance with the manufacturer's instructions.
Romberg N., Virdee M., Chamberlain N., Oe T., Schickel J.N., Perkins T., Cantaert T., Rachid R., Rosengren S., Palazzo R., Geha R., Cunningham-Rundles C, & Meffre E. (2015). TNFRSF13B hemizygosity reveals TACI haploinsufficiency at later stages of B-cell development. The Journal of allergy and clinical immunology, 136(5), 1315-1325.
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2

Comprehensive Immune Cell Phenotyping in Pancreatitis

Cited 1 time
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The following antibodies were used for FACS staining, IHC staining and immunofluorescence staining: Anti-mouse CD4 BV650 (BioLegend 100546), anti-mouse-CD11b PerCP Cy55 (BioLegend, 101228), anti-mouse-Ly6G BV421 (BioLegend, 127628), anti-mouse-Ly6C BV605 (BioLegend 128036), anti-CD25-PECy7 (BioLegend, 102016), anti-CD69 BV510 (BioLegend, 104532), anti-CD8a BV605 (BioLegend, 100743), anti-FoxP3 APC (Miltenyi Biotec, 130-111-601), anti-mouse-CD11b (abcam, Ab133357), anti-mouse-CD68 (antibody-online, ABIN181836), anti-CCR2 (abcam, ab273050) anti-Ki67 (Bethyl, IHC-00375), anti-mouse-CD11b (abcam, Ab133357), anti-mouse-Ly6g (abcam, Ab25377), anti-mouse-Cystatin C (Novus biologicals, NB100-1033), anti-VE-cadherin (abcam, ab7047-50) anti-rabbit-HRP (DAKO, K4003), anti-mouse-HRP (DAKO, K4001). Caerulein was obtained from Sigma Aldricht (C9026-1MG, Munich, Germany), human myeloperoxidase from Calbiochem (Cat# 475911). All antibodies used in this study had been tested and established in previous studies (11 (link), 12 (link), 15 (link)). The concentration of the antibodies was used according to the manufacturer’s instructions.
Wilden A., Glaubitz J., Otto O., Biedenweg D., Nauck M., Mack M., Ribback S., Bröker B.M., von Rheinbaben S.F., Lerch M.M., Aghdassi A.A., Weiss F.U, & Sendler M. (2022). Mobilization of CD11b+/Ly6chi monocytes causes multi organ dysfunction syndrome in acute pancreatitis. Frontiers in Immunology, 13, 991295.
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3

Multiparametric Flow Cytometry of T-Cell Subsets

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T-cell subsets were enumerated in freshly thawed cryopreserved PBMC. After washing and counting viable cells, PBMCs were surface-stained with the following conjugated mAbs: anti-CD3-AF488 (Biolegend; clone HIT3a), anti CD4-APC/Cy7 (Biolegend; RPA-T4), anti-CD25-PE/Cy7 (Biolegend; BC96), anti-HLA-DR-PerCP/Cy5.5 (Biolegend; L243), anti-CD39-APC (Biolegend; A1), and anti-CD38-PECy7 (Biolegend; HIT2). Cells were fixed and permeabilized with Cytofix/Cytoperm (BD Biosciences), and stained with anti-IL10-APC (R & D Systems; 127107), anti-FOXP3-PE (Biolegend; 206D), anti-TGFβ-PerCP/Cy5.5 (Biolegend; TW4-2F8) and anti-IL35-PE (eBioscience; ebic6) and analyzed with Guava easyCyte 8HT and FlowJo (Treestar).
Subsets were expressed as percentages of the parent CD4+ and CD8+ T-cell populations. The gating strategy is presented in Fig S1.
Weinberg A., Park J.G., Bosch R., Cho A., Livingston E., Aweeka F., Cramer Y., Watts D.H., Luque A.E, & Cohn S.E. (2016). EFFECT OF DEPOT MEDOXYPROGESTERONE ACETATE ON IMMUNE FUNCTIONS AND INFLAMMATORY MARKERS OF HIV-INFECTED WOMEN. Journal of acquired immune deficiency syndromes (1999), 71(2), 137-145.
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4

Phenotypic Characterization of NY-ESO-1 T Cells

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To exclude dead cells, the Near-IR Dead Cell Stain Kit (Thermo Fisher Scientific) was used. NY-ESO-1-specific TCR expression was evaluated using the PE-conjugated A0201 NY-ESO-1-tetramer (kindly provided by Hiroshi Shiku, Mie University, Tsu, Japan). The following fluorochrome-conjugated antibodies were used for immunophenotyping of different surface markers on SW982 cells as well as NY-ESO-1-specific T cells: anti-CD80-APC (BioLegend, San Diego, CA, USA), anti-HLA-ABC-FITC (eBioscience, San Diego, CA, USA), anti-programmed cell death protein ligand 1 (PD-L1)-PE (eBioscience), anti-CD3-V500 (AmCyan) (BD Biosciences, San Diego, CA, USA), anti-CD8-Parcific blue (BioLegend), anti-CD25-PE-Cy7 (BioLegend), anti-CD4-Alexa Fluor 700 (eBioscience). Data acquisition was performed on an LSR II device (BD Biosciences) and data were analyzed using the FlowJo software (TreeStar, Ashland, OR, USA).
Gong W., Wang L., Schubert M.L., Kleist C., Neuber B., Wang S., Yang M., Hückelhoven-Krauss A., Wu D., Schmitt A., Müller-Tidow C., Shiku H., Schmitt M, & Sellner L. (2022). HDAC Inhibition for Optimized Cellular Immunotherapy of NY-ESO-1-Positive Soft Tissue Sarcoma. Biomedicines, 10(2), 373.
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5

Comprehensive Immune Cell Profiling

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T-cell activation/exhaustion was analyzed by staining with anti-CD3-PerCP, anti-CD4-PacificBlue, anti-CD8-APC, anti-CD25-PE/Cy7 and anti-CD279-PE (PD-1); ILCs by staining with anti-Lineage-Cocktail-APC (anti-CD3/CD14/CD16/CD19/CD20/CD56) and anti-CD127-PE; NK-cell subsets by staining with anti-CD3-PerCP, anti-CD14-FITC, anti-CD19-PE/Cy7, anti-CD56-APC/Cy7, anti-CD16-PacificBlue (all BioLegend, Fell, Germany), anti-NKG2A-APC (clone 131411) and anti-NKG2C-PE (both R&D Systems, Abingdon, UK) [20 (link)]. Samples were acquired on a CyAn ADP Analyzer (Beckman Coulter, Nyon, Switzerland) and data analyzed with FlowJo software vX.0.7 (FlowJo, Ashland, Oregon, USA).
Stuehler C., Bernardini C., Elzi L., Stoeckle M., Zimmerli S., Furrer H., Günthard H.F., Leibundgut-Landmann S., Battegay M, & Khanna N. (2016). Immune recovery in HIV-infected patients after Candida esophagitis is impaired despite long-term antiretroviral therapy. AIDS (London, England), 30(12), 1923-1933.
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6

Flow Cytometric Immunophenotyping of Lymphocytes

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The following antibodies were used for flow cytometric stainings: anti-CD19 APC-Cy7, anti-CD27 PerCP-Cy5.5, anti-CD21 Pacific Blue, anti-CD4 APC-Cy7, anti-CD25 PE- Cy7, PE or PE Dazzle, anti-CD127 PerCP-Cy5.5 or APC, anti-CD45RO AF-700, anti-CXCR5 Pacific Blue, anti-PD-1 PE-Cy7 (all from BioLegend), anti-CD3 eFluor 605NC (eBioscience) and anti-IgG APC (Becton Dickinson). Intracellular staining with anti-Foxp3 Alexa Fluor 488 (eBioscience) or APC (Biolegend) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) in accordance with manufacturer instructions.
Romberg N., Le Coz C., Glauzy S., Schickel J.N., Trofa M., Nolan B.E., Paessler M., Xu M.L., Lambert M., Lakhani S.A., Khokha M.K., Jyonouchi S., Heimall J., Takach P., Maglione P.J., Catanzaro J., Hsu F.I., Sullivan K.E., Cunningham-Rundles C, & Meffre E. (2018). CVID patients with autoimmune cytopenias exhibit hyperplastic yet inefficient germinal center responses. The Journal of allergy and clinical immunology, 143(1), 258-265.
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7

Characterization of B and T Cell Subsets in Systemic Sclerosis

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Mononuclear cells from healthy donors and patients with SSc were enriched for B cells by magnetic separation with anti-CD20 microbeads (Miltenyi Biotech) and stained with anti-human CD19-Pacific Blue, anti-human CD27-PerCP Cy5.5, anti-human CD10-PE-Cy7, anti-human CD21-APC, anti-human IgM-FITC (Biolegend). Single CD19+CD21lowCD10+IgMhiCD27− new emigrant/transitional and CD19+CD21+CD10−IgM+CD27− mature naïve B cells were sorted on a FACSAria (BD Biosciences) into 96-well PCR plates and immediately frozen on dry ice.
The following antibodies were used for T cell phenotyping: anti-CD4 APC-Cy7, anti-CD25 PE-Cy7, anti-CD127 PerCP-Cy5.5 (all from Biolegend), and anti-CD3 eFluor 605NC (eBioscience). Intracellular staining with anti-FOXP3 Alexa Fluor 488 (clone PCH101; eBioscience) was performed using the FOXP3/Transcription Factor Staining Buffer Set in accordance with the manufacturer’s instructions (eBioscience).
Glauzy S., Olson B., May C.K., Parisi D., Massad C., Hansen J.E., Ryu C., Herzog E.L, & Meffre E. (2021). Defective early B cell tolerance checkpoints in patients with systemic sclerosis allow the production of self-antigen-specific clones. Arthritis & rheumatology (Hoboken, N.J.), 74(2), 307-317.
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8

Tumor Immune Cell Profiling by Flow Cytometry

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Tumor immune cells isolated as described above were resuspended at 106 cells/100 μl in FACS buffer (2% BSA, 2% goat serum in PBS). Cells were immunolabeled with the following fluorochrome-conjugated cell surface antibodies: anti-CD45 PE/Cy5 (0.25 μg/100 μl), anti-CD11b PerCP (0.25 μg/100 μl), anti-CD4 FITC (0.25 μg/100 μl), anti-CD8 PE (0.25 μg/100 μl), anti-CD25 PE/Cy7 (0.5 μg/100 μl), and anti-FOXP3 Alexa Fluor 647 (5 μl/100 μl) all purchased from BioLegend. Single cells were prepared for flow cytometry as previously described [23 ] or according to the manufacturer’s protocol (BioLegend, for FOXP3). Cell-associated fluorescence was acquired and analyzed using the BD LSR II cytometer and TreeStar Inc. FlowJo software, respectively.
Obr A.E., Kumar S., Chang Y.J., Bulatowicz J.J., Barnes B.J., Birge R.B., Lazzarino D.A., Gallagher E., LeRoith D, & Wood T.L. (2018). Insulin-like growth factor receptor signaling in breast tumor epithelium protects cells from endoplasmic reticulum stress and regulates the tumor microenvironment. Breast Cancer Research : BCR, 20, 138.
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9

Effector Treg Cell Isolation Protocol

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To sort the effector Treg cells, the following monoclonal antibodies were used: anti-CD3 (APC; BD Bioscience, USA), anti-CD4 (PerCP cy5.5; BD Bioscience), anti-CD45RA (APC cy7; BioLegend, USA), anti-CD25 (PE cy7; BioLegend), and anti-CD127 (PE; BD Bioscience). PBMC and decidual mononuclear cells were stained by these antibodies for 20 min on ice. After staining, the cells were washed with PBS and analyzed using a FACSAria II flow cytometer (BD Biosciences). CD3+CD4+CD45RA−CD25+CD127low/− effector Treg cells were single-cell sorted into wells of a 96-well PCR plate. The gating strategy used to sort the effector Treg cells is presented in Figure 1.
Tsuda S., Zhang X., Hamana H., Shima T., Ushijima A., Tsuda K., Muraguchi A., Kishi H, & Saito S. (2018). Clonally Expanded Decidual Effector Regulatory T Cells Increase in Late Gestation of Normal Pregnancy, but Not in Preeclampsia, in Humans. Frontiers in Immunology, 9, 1934.
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10

Comprehensive Immune Cell Profiling

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B- and T-cell subsets were enumerated in freshly thawed cryopreserved PBMCs. After washing and counting viable cells, PBMCs were surface-stained with the following conjugated mAbs: anti-CD3-AF488 (Biolegend; clone HIT3a), anti CD8-APC/Cy7 (Biolegend; SK1), anti-CD19-APC/Cy7 (BD Biosciences; SJ25C1), anti-CD19-PerCP/Cy5.5 (Biolegend; HiB19) anti-CD25-PE/Cy7 (Biolegend; BC96), anti-CD21-PE/Cy7 (BD Biosciences; B-ly4), anti-CD27-PerCP/Cy5.5 (BD Biosciences; M-T271), anti-CD38-PE/Cy7 (Biolegend; HIT2), anti-HLA-DR-PerCP/Cy5.5 (Biolegend; L243); anti-CD10-FITC (BD Biosciences; W8E7), anti-IL21R-PE (BD Biosciences; 17A12), and anti-CD39-APC (Biolegend; A1). Cells were fixed and permeabilized with Cytofix/Cytoperm (BD Biosciences), and stained with anti-IL-10-PE (R & D Systems; 127107), anti-FOXP3-PE (Biolegend; 206D) and anti- TGFβ-PE (Biolegend; TW4-2F8), and analyzed with Guava easyCyte 8HT and FlowJo (Treestar). Subsets were expressed as percentages of the parent CD4+, CD8+ and CD19+ cell populations. The gating strategy is presented in S2 Fig.
Weinberg A., Muresan P., Richardson K.M., Fenton T., Dominguez T., Bloom A., Watts D.H., Abzug M.J., Nachman S.A, & Levin M.J. (2015). Determinants of Vaccine Immunogenicity in HIV-Infected Pregnant Women: Analysis of B and T Cell Responses to Pandemic H1N1 Monovalent Vaccine. PLoS ONE, 10(4), e0122431.
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