Anti foxp3 clone 206d

Manufactured by BioLegend

Anti-FoxP3 (clone 206D) is a mouse monoclonal antibody that binds to the transcription factor FoxP3, which is a critical regulator of regulatory T cell (Treg) development and function. This antibody can be used for the identification and analysis of FoxP3-expressing cells in various applications such as flow cytometry.

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Anti-Foxp3 (35 citations) Clone 206D (6 citations) FoxP3 clone 206D (5 citations) Anti-Foxp3 (206D) (2 citations) PE-conjugated anti-human Foxp3 (clone 206D), (2 citations)

2 protocols using anti foxp3 clone 206d

Expansion and Purification of Human Regulatory T Cells

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Freshly isolated, blood-derived T cell populations were cultured in 96-well round bottom plates (Costar) coated with 0.5 ug/ml anti-CD3 (UCHT-1, eBioscience) with irradiated antigen presenting cells (iAPC, PBMCs depleted of T cells via anti-CD2-beads from Dynal/Invitrogen) and 0.25 ug/ml anti-CD28 (clone 28.2 eBioscience) in RPMI-1640 medium (Life Technologies) supplemented with 10% FBS (Life Technologies) and 20 U/ml rhIL-2. Confluent wells were split into new plates and IL-2 containing media was supplemented every 2–3 days. After two weeks, purity (expression of FoxP3^high) was tested by flow cytometry using anti-FoxP3 (clone 206D, BioLegend) and the intracellular FoxP3 staining kit (Biolegend).

Donnelly C., Dykstra B., Mondal N., Huang J., Kaskow B.J., Griffin R., Sackstein R, & Baecher-Allan C. (2018). Optimizing human Treg immunotherapy by Treg subset selection and E-selectin ligand expression. Scientific Reports, 8, 420.

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Multiparametric Immunophenotyping of PBMCs

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Whole blood samples were collected in cell preparation tubes with sodium citrate (BD Biosciences). PBMCs were obtained by centrifugation and viably frozen until analysis. PBMCs were thawed, washed with flow buffer (5% BSA, 2 mM EDTA in PBS) and incubated with LIVE/DEAD Fixable Aqua Dead Cell Stain (Life Technologies), Fc receptor blocking agent (Miltenyi Biotec) and stained with surface antibodies (CD3 clone OKT3, CD4 clone RPA-T4, CD8 clone SK1, CD14 clone HCD14, CD19 clone HIB19, CD25 clone BC96, ICOS clone C398.4A, HLA-DR clone L243, PD-1 clone 29F.1A12 all from BioLegend) for 20 min at 4°C. For Foxp3 and Ki67 staining, cells were fixed and permeabilized using a Fix/Perm buffer (eBiosciences) according to the manufacturer’s instructions, then stained with anti-Foxp3 (clone 206D, BioLegend) or anti-Ki67 antibody (clone B56, BD Biosciences).
For global protein acetylation analysis, after surface staining, cells were fixed with 0.4% paraformaldehyde (Thermo Fisher Scientific), permeabilized in Triton X-100 (Sigma-Aldrich) and subsequently stained with anti-acetylated lysine antibody (clone 15G10, BioLegend).

Hellmann M.D., Jänne P.A., Opyrchal M., Hafez N., Raez L.E., Gabrilovich D., Wang F., Trepel J.B., Lee M.J., Yuno A., Lee S., Brouwer S., Sankoh S., Wang L., Tamang D., Schmidt E., Meyers M.L., Ramalingam S.S., Shum E, & Ordentlich P. (2020). Entinostat plus pembrolizumab in patients with metastatic NSCLC previously treated with anti-PD-(L)1 therapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(4), 1019-1028.

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