Foxp3 clone 236a e7

Manufactured by BD

Sourced in United States

FOXP3 (Clone 236A/E7) is a mouse monoclonal antibody that recognizes the FOXP3 protein. FOXP3 is a transcription factor that plays a critical role in the development and function of regulatory T cells (Tregs). The antibody can be used for the identification and characterization of FOXP3-expressing cells in various applications.

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3 protocols using foxp3 clone 236a e7

T-Cell Isolation and Phenotyping

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Prior to and following magnetic bead isolation procedures, T cells were analyzed using flow cytometry to assess the efficiency of T-cell isolation procedures. To permit T cell phenotyping, cells were washed and then stained for surface expression of CD4 (BD Pharmingen, San Diego, USA), CD25 (BD Biosciences, San Jose, USA) and CD127 (clone hIL-7R-M21) (BD Pharmingen, San Diego, USA) followed by intra-cellular staining for FOXP3 (Clone 236A/E7) (BD Pharmingen, San Diego, USA). Flow cytometry was performed using an LSR II flow cytometer (BD Biosciences, San Jose, USA). Cytobank software was used to analyze data generated from flow cytometry. Following bead enrichment, cells were harvested and washed twice with ice-cold PBS, snap-frozen in liquid nitrogen and then stored at −80 until further sample preparation.

Olsson N., Schultz L.M., Zhang L., Khodadoust M.S., Narayan R., Czerwinski D.K., Levy R, & Elias J.E. (2018). T-cell immunopeptidomes reveal cell subtype surface markers derived from intracellular proteins. Proteomics, 18(12), e1700410.

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Comprehensive Immune Cell Profiling

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PBMCs were stained with fluorochrome-conjugated murine monoclonal antibodies against human CD3 (clone HIT3a), CD4 (clone RPA-T4), CD8 (clone RPA-T8), CD45RA (clone HIT100), CD45RO (clone UCHL1), CD25 (clone M-A251), CD127 (clone HIL-7R-M21), CD19 (clone HIB19), CD27 (clone M-T271), CD38 (clone HIT2), IgD (clone IA6-2), CD24 (clone ML5) and FoxP3 (clone 236A/E7) (all from BD Biosciences, San Jose, CA, USA). Multicolor flow cytometry was used to identify T- and B-cell subsets with standard technique and equipment (FACS FortessaX20, BD Biosciences and Flow-jo software, version 10.7.1).

Casiraghi F., Todeschini M., Podestà M.A., Mister M., Ruggiero B., Trillini M., Carrara C., Diadei O., Villa A., Benigni A, & Remuzzi G. (2023). Immunophenotypic Alterations in Adult Patients with Steroid-Dependent and Frequently Relapsing Nephrotic Syndrome. International Journal of Molecular Sciences, 24(9), 7687.

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Comprehensive Treg Immunophenotyping and Epigenetics

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In vitro-expanded Tregs were stained with Live/Dead Aqua (Life Technologies) followed by the following anti-human antibodies; CD4 BUV496 (clone SK3, BD), CD127 APC-Vio770 (clone REA614, Miltenyi), CD25 APC (clone BC96, Biolegend) and TCR Vbeta21.3 PE (clone REA894, Miltenyi).
For intracellular staining: Tregs were stimulated with PMA/ionomycin and Brefeldin A (Sigma) for 4 h 37 °C after which cells were fixed and permeabilized with Foxp3/Transcription Factor Staining Buffer Set (eBioscience) and stained with anti-human antibodies; Foxp3 (clone 236A/E7, BD), Helios (clone 22F6, Biolegend), IFN-γ (clone B27, BD) and IL-17A (clone N49-653, BD). Cells were acquired on a Cytek Aurora 5 L flow cytometer and analysis performed in FlowJo (version 10.6.2). A gating strategy is found in Supplementary Fig. 7. Methylation of the FOXP3 TDSR was performed by pyrosequencing of intron 1 of TDSR region -2330 to -2263 from ATG of FOXP3 and analysis of 9 CpG sites by EpigenDx.

Eggenhuizen P.J., Cheong R.M., Lo C., Chang J., Ng B.H., Ting Y.T., Monk J.A., Loh K.L., Broury A., Tay E.S., Shen C., Zhong Y., Lim S., Chung J.X., Kandane-Rathnayake R., Koelmeyer R., Hoi A., Chaudhry A., Manzanillo P., Snelgrove S.L., Morand E.F, & Ooi J.D. (2024). Smith-specific regulatory T cells halt the progression of lupus nephritis. Nature Communications, 15, 899.

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