Foxp3 259d c7

Manufactured by BD

Sourced in United Kingdom

Foxp3 (259D/C7) is a lab equipment product used for the detection and analysis of the Forkhead box protein P3 (Foxp3), which is a key transcription factor involved in the regulation of T-cell development and function. The product is designed to provide accurate and reliable results in research and clinical applications.

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3 protocols using foxp3 259d c7

Murine Melanoma Immunotherapy Protocols

Cited 1 time

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The FBL cell line was a gift from Dr. Philip Greenberg (University of Washington) in 2008 and has been described previously (20 , 21 (link)). FBL has not been authenticated. The FBL cell line is maintained in vivo and cells are harvested from ascites fluid on the day of experiment setup. The HLA-A2⁺ human melanoma line MeWo was purchased from ATCC in 2014. Peptides from FBL-Gag (CCLCLTVFL) and ovalbumin (SIINFEKL) were obtained from GenScript. Mouse blocking antibodies to CTLA-4 (9D9), PD-1 (RMP1–14) and LAG-3 (C9B7W) were purchased from BioXCell. Human antibodies against CTLA-4, PD-1, and LAG-3 were provided by Bristol-Myers Squibb. All blocking antibodies were administered intraperitoneally (i.p.) at a dose of 100 µg/mouse every 3 days. Fluorochome-conjugated antibodies to mouse CD90.1 (OX-7), CD90.2 (53–2.1), IFNγ (XMG1.2), TNF (MP6-XT22), and anti-CD16/CD32 F_c block (2.4G2) and antibodies to human CD45 (HI30), CD3 (UCHT1), CD4 (RPA-T4), CD8 (SK1), and Foxp3 (259D/C7) were purchased from BD Biosciences. Fluorochrome-conjugated antibody to CD8 (53–6.7) was purchased from BioLegend. Fluorochrome-conjugated antibodies to mouse CD4 (GK1.5), NK1.1 (PK136), Eomes (Dan11mag), and Foxp3 (FJK-16s) and antibody to human T-bet (ebio4b10) were purchased from eBioscience.

Klevorn L.E., Berrien-Elliott M.M., Yuan J., Kuehm L.M., Felock G.D., Crowe S.A, & Teague R.M. (2016). Rescue of tolerant CD8+ T cells during cancer immunotherapy with IL2:antibody complexes. Cancer immunology research, 4(12), 1016-1026.

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Immunohistochemical Analysis of Tumor Markers

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Immunohistochemistry was performed on 2‐μm FFPE sections in an automated immunostainer (Autostainer Link48; Dako, Glostrup, Denmark) according to the manufacturer's instructions. The following antibodies were used: β‐catenin 14 (1 : 1000; BD Transduction, South San Francisco, CA, USA), CD3 polyclonal (1 : 400; Dako), CD4 4B12 (1 : 300; Dako), CD8 C8/144B (1 : 20; Dako), CD20 L26 (1 : 400; Dako), CD21 1F8 (1 : 50; Dako), CD56 123C3 (1 : 400; Dako), CD68 KP1 (1 : 3000; Dako), CD163 10D6 (1 : 200; Novocastra, Newcastle, UK), FOXP3 259D/C7 (1 : 200; BD Pharmingen), PD‐L1 SP142 (1 : 30; SPRING, Pleasanton, CA, USA), PD1 NAT105 (1 : 100; Biocare Medical, Pikenine, Concord, NC, USA), and EZH2 5246S (1 : 50; Cell Signaling, Leiden, Netherlands).

Colombo C., Belfiore A., Paielli N., De Cecco L., Canevari S., Laurini E., Fermeglia M., Pricl S., Verderio P., Bottelli S., Fiore M., Stacchiotti S., Palassini E., Gronchi A., Pilotti S, & Perrone F. (2017). β‐Catenin in desmoid‐type fibromatosis: deep insights into the role of T41A and S45F mutations on protein structure and gene expression. Molecular Oncology, 11(11), 1495-1507.

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Multiparametric Flow Cytometry Analysis

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The following antibodies were used for flowcytometry: CD3 (HIT3a, Becton-Dickinson (BD)), CD4 (SK3, BD), CD45RA (HI100, Biolegend), CXCR5 (TG2/CXCR5,Biolegend and RF8B2,BD), CCR7 (3D12, BD), PD-1 (NAT), CD25 (2A3, BD), CD127 (A7R34, eBioscience), CRTAM(R&D), Granzyme A (GB11,BD), Foxp3 (259D/C7, BD), CD57(HNK-1,BD), IL-17 (SCPL1362, BD), IL-10 (JES3-19F1,BD), IFN-g (25723.11,BD), IL-4 (7A3-3,BD), IL-21 (3A3-N2.1, BD), STAT3(4/P-STAT3,BD), and cell trace violet (CTV,Life Technologies). Data were acquired on FACSCaliber or FACSCanto II and data were then analyzed with FlowJo, version (8.7) (Treestar).

Alshekaili J., Chand R., Lee C.E., Corley S., Kwong K., Papa I., Fulcher D.A., Randall K.L., Leiding J.W., Ma C.S., Wilkins M.R., Uzel G., Goodnow C.C., Vinuesa C.G., Tangye S.G, & Cook M.C. (2018). STAT3 regulates cytotoxicity of human CD57+ CD4+ T cells in blood and lymphoid follicles. Scientific Reports, 8, 3529.

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