Foxp3 pe

Manufactured by Beckman Coulter

Sourced in United States

The FOXP3-PE is a fluorochrome-conjugated antibody that binds to the Forkhead box protein P3 (FOXP3), a transcription factor expressed in regulatory T cells. This product is suitable for the identification and characterization of FOXP3-expressing cells by flow cytometry.

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Lab products found in correlation

4 protocols using foxp3 pe

Multi-Color Flow Cytometry Immunophenotyping

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Antibody combination panels for four or five color immune-staining were as follows:

CD3PerCP, CD4 FITC, CD25APC, and rat Foxp3PE (all from eBioscience, San Diego, CA).

CD4 PercpCy5.5, HLA-DR APC, CD127 Pac Blue and rat Foxp3 PE (eBioscience, San Diego, CA).

CD4 PercpCy5.5, CCR5 FITC, CXCR4 APC (Biolegend, San Diego, CA) and Foxp3 PE.

CD4 PercpCy5.5, HIV-1 p24 FITC (Beckman Coulter, Brea, CA), Bcl-2 PE (BD Biosciences, San Jose, CA) and Foxp3 e-fluor660 (eBioscience).

CD4 PercpCy5.5, CD127 Pac Blue, Foxp3 PE, HIV-1 p24 FITC and IFN-γ Alexa Fluor 647 (Biolegend).

Isotype antibodies used included: IgG2a APC, IgG1 Pac Blue, rat IgG2a PE, IgG2a FITC, rat IgG2a AP, IgG1 FITC, IgG1 PE, IgG2a e-fluor660, and IgG1 Alexa Fluor 647.
The protocol and buffer set from eBioscience were used for all experiments involving intracellular staining. All samples were fixed and acquired within 1h of completion of staining, and analysis was by Miltenyi MACSQuant Flow cytometer.
Data were analyzed by FlowJo software (Treestar Inc, Ashland, OR) at the completion of study.

Hirsch C.S., Baseke J., Kafuluma J.L., Nserko M., Mayanja-Kizza H, & Toossi Z. (2016). Expansion and productive HIV-1 infection of Foxp3 positive CD4 T cells at pleural sites of HIV/TB co-infection. Journal of clinical & experimental immunology, 1(1), http://www.opastonline.com/wp-content/uploads/2016/11/expansion-and-productive-hiv-1-infection-of-foxp3-positive-cd4-t-cells-at-pleural-sites-of-hivtb-co-infection-jcei-16-007.pdf.

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Regulatory T Cell Phenotyping by Flow Cytometry

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For the analysis of Tregs, non-adherent PBMCs were co-cultured, for 7 days with TLDCs (1:10 ratio), in the presence or absence of cisplatin as done for the proliferation assay, but without CFSE staining. Anti- CD4-PC5, CD8-APC, CD25-ECD (Cat. No.: 6607112, clone: B1.49.9), CD3-PC7 (Cat. No.: 6607100, clone:259D) (all from Beckman Coulter Inc.) and FOXP3-PE (Cat. No.: 320108, clone:206D from BioLegend) markers were used to stain the cells after a week for Treg analysis by flow cytometry. A fixable live dead discriminator (Thermo Fisher Scientific) was used to estimate the percentage of live/dead cells.

Dhandapani H., Seetharaman A., Jayakumar H., Ganeshrajah S., Singh S.S., Thangarajan R, & Ramanathan P. (2021). Autologous cervical tumor lysate pulsed dendritic cell stimulation followed by cisplatin treatment abrogates FOXP3+ cells in vitro. Journal of Gynecologic Oncology, 32(4), e59.

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Phenotyping of NKT cells and Tregs

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NKT-PBMCs and whole peripheral blood samples from healthy controls and CTD-IP patients were stained with the following antibodies: CD3-FITC, CD56-PE, CD127-PE, CD45-ECD, CD4-FITC, CD25-PC5, CD4-FITC/CD8-PE/CD3-PC5, FOXP3–PE, and appropriate isotype controls (Beckman Coulter, Indianapolis, IN, USA). Staining was performed according to the manufacturer’s instructions.

Liu M., Zeng X., Wang J., Fu Z., Wang J., Liu M., Ren D., Yu B., Zheng L., Hu X., Shi W, & Xu J. (2016). Immunomodulation by mesenchymal stem cells in treating human autoimmune disease-associated lung fibrosis. Stem Cell Research & Therapy, 7, 63.

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Flow Cytometry Analysis of T Cell Subsets

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Flow cytometry: T cell subsets were identified using multicolor flow cytometry with standard techniques on the Navios EX flow cytometer (Beckman Coulter, Sykesville, MD, United States). Whole blood samples were drawn from patients at the scheduled time points (T0, T3, and T6), collected in EDTA tubes, and processed for the evaluation of lymphocyte count and their subpopulations. T lymphocyte subsets determined were CD3+CD4+, CD3+CD8+, CD3-CD16+CD56+, CD3+CD4+CD28-, and CD3+CD8+CD28-, using the following monoclonal antibodies: CD3-FITC (clone: UCHT1; Beckman Coulter), CD16 (clone: 3G8; Beckman Coulter), CD56 clone: N901(NKH-1)-PE; Beckman Coulter), CD4-APC (clone: 13B8.2; Beckman Coulter), CD8 PC5.5 (clone: B9.11l Beckman Coulter), CD28-ECD (clone: CD28.2; Beckman Coulter), and CD45-PC7 (clone: J33; Beckman Coulter). Peripheral blood mononuclear cells were obtained by Ficoll density gradient centrifugation. Immunophenotyping of Tregs was performed with the combination of the following monoclonal antibodies: CD45-PC7 (clone: J33; Beckman Coulter), CD4-FITC (clone: 13B8.2; Beckman Coulter), CD25-PC5 (clone: B1.49.9; Beckman Coulter), and FOXP3-PE (clone: 259D; Beckman Coulter).

Vagiotas L., Stangou M., Kasimatis E., Xochelli A., Myserlis G., Lioulios G., Nikolaidou V., Panteli M., Ouranos K., Antoniadis N., Maria D., Papagianni A., Tsoulfas G, & Fylaktou A. (2022). Effect of panel reactive antibodies on T cell immunity reinstatement following renal transplantation. World Journal of Transplantation, 12(10), 313-324.

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