Anti foxp3 pe fjk 16s

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-Foxp3–PE (FJK-16s) is a fluorochrome-conjugated antibody specific for the Foxp3 transcription factor. Foxp3 is a key regulator of the development and function of regulatory T cells (Tregs). This antibody can be used to detect and quantify Foxp3-expressing cells by flow cytometry.

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Lab products found in correlation

Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (5 mentions) Anti il 4 pe 11b11, by BD (4 mentions) Anti il 5 pe trfk5, by BD (4 mentions) Anti cd25 apc pc61, by Thermo Fisher Scientific (4 mentions) Anti ifn γ pe or per cp 5.5 xmg1 2, by BD (3 mentions) Facsdiva software, by BD (3 mentions) Anti cd3 pe cy7 145 2c11, by Thermo Fisher Scientific (2 mentions) Lsrii facs analyzer, by BD (1 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions) Anti cd4 pe cy7, by BD (1 mentions) Lsrfortessa x 20, by BD (1 mentions) Anti ifn γ apc, by BD (1 mentions) Anti il 17a pe, by BD (1 mentions) Fitc anti gfp, by Abcam (1 mentions) Macsquant, by Miltenyi Biotec (1 mentions) Facscanto 2, by BD (1 mentions) Permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Anti cd4 pe l3t4 gk1.5, by BD (1 mentions) Facsaria cell sorter, by BD (1 mentions)

Spelling variants of this product

FJK-16s (166 citations) Anti-Foxp3 (119 citations) Foxp3-PE (109 citations) Foxp3 (FJK-16s, (98 citations) Anti-Foxp3-PE (90 citations) Anti-Foxp3 (FJK-16s) (63 citations) Foxp3 PE (FJK-16s) (6 citations) Anti-Foxp3-PE (clone FJK-16s; (6 citations) PE)-Cy5-conjugated anti-mouse Foxp3 (FJK-16s) (2 citations) Anti-mouse Foxp3-PE (clone FJK-16S) (2 citations)

17 protocols using anti foxp3 pe fjk 16s

Comprehensive Multicolor Flow Cytometry

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Cells were stained for surface markers with the following antibodies: Pacific-blue- or Alexa-488-anti-CD4 (RM4-5, GK1.5); Pacific-blue-anti-CD19 (6D5); PE conjugated anti-PD-1 (RMP1-30), CD44 (IM7), TGF-β1 (TW7-16B4) all from Biolegend. Alexa-647-anti-GL-7 (GL7); FITC- or PE-anti-ICOS (398.4A or 7E.17G9); PE conjugated anti-CD25 (pc61.5), GITR (DTA-1), and Fas (15A7) all from eBioscience; PE-Cy7-anti-CXCR5 (2G8, BD Biosciences); PE-anti-CTLA-4 (UC10-4F10-11, BD Pharmingen).
For nuclear transcription factor staining, cells were labeled with surface markers, then fixed and permeabilized with the Foxp3-Staining-Buffer-Set (eBioscience), according to the manufacturer's instruction. Cells were then stained with PE-anti-Bcl6 (K112-91, BD Biosciences) and PE-anti-Foxp3 (FJK-16s, eBiosciences).
For phospho-flow staining, after treatment, cells were fixed and permeabilized with the BD Phosflow™ Fix Buffer and Perm Buffer, according to the manufacturer's instruction. Surface markers staining were followed by intracellular staining with Alexa-647-rabbit-anti-phospho-Akt-Ser473 (Cell signaling) or Pacific-blue-mouse-anti-Stat3-p-Y705 (4/p-Stat3, BD Bioscience). Samples were acquired with an LSRII FACS analyzer (BD Biosciences), and data was analyzed with FlowJo software (Tree Star, Inc. Ashland, OR, USA).

Ding Y., Li J., Yang P., Luo B., Wu Q., Zajac A.J., Wildner O., Hsu H.C, & Mountz J.D. (2014). Interleukin-21 Promotes Germinal Center Reaction by Skewing the Follicular Regulatory T Cell to Follicular Helper T Cell Balance in Autoimmune BXD2 Mice. Arthritis & rheumatology (Hoboken, N.J.), 66(9), 2601-2612.

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Comprehensive Antibody Panel for Immune Cell Analysis

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The following antibodies were purchased from BioLegend: PerCP-Cy5.5 anti-B220 (RA3-6B2), PE anti-CD103 (2E7), APC-Cy7 anti-CD11c (N418), PE-Cy7 anti-CD4 (RM4-5), BV510 anti-CD45 (30-F11), PerCP-Cy5.5 anti-CD8a (53-6.7), APC anti-F4/80 (BM8), APC anti-CXCR5 (L138D7), Brilliant violet 421 anti-PD1 (29F.1A12), FITC anti-IgMb (AF6-78), PE anti-IgMa (MA-69), and FITC anti-Vβ8.1/8.2 (KJ16-133.18). The following antibodies were purchased from eBioscience: Alexa Fluor 488 anti-GL7 (GL7), PE-Cy7 anti-IgD (11-26), PerCP-eFluor 710 anti-Fas (15A7), PE or eFluor 450 anti-CD19 (1D3), APC anti-CD25 (PC61.5), FITC anti-CD44 (IM7), and PE anti-FoxP3 (FJK-16s). The following antibodies were purchased from BD: PE anti-CD62 ligand (MEL-14), FITC or APC anti-CD3e (145-2C11), and FITC anti-Vβ4 (KT4). APC-conjugated F(ab′)₂ donkey anti–mouse IgG (H+L) was purchased from Jackson ImmunoResearch Laboratories, Inc., and Pacific blue I-A^g7 (AG2.42.7) was made in our laboratory.

Wan X., Thomas J.W, & Unanue E.R. (2016). Class-switched anti-insulin antibodies originate from unconventional antigen presentation in multiple lymphoid sites. The Journal of Experimental Medicine, 213(6), 967-978.

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Antibody Staining for T-Cell Analysis

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PC61 (anti-CD25) was purchased from Bioexpress (NH) for in vivo administration. Antibodies purchased from BD Pharmingen were PerCP anti-CD8α (clone; 53-6.7), Alexa Flour 700 anti-IFN-γ (clone; XMG1.2), APC anti-IL-2 (Clone; JHS6-5H4), and PE-Cy7 anti-CD4 (clone; GK1.5). Antibodies purchased from eBioscience were FITC anti-CD4 (RM4-5), APC anti-CD25 (eBio,7D4), and PE anti-Foxp3 FJK-16s.

Qin L., Jiang G., Han J, & Letvin N.L. (2015). Regulatory T Cells Modulate DNA Vaccine Immunogenicity at Early Time via Functional CD4+ T Cells and Antigen Duration. Frontiers in Immunology, 6, 510.

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Flow Cytometry for Immune Cell Analysis

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Mononuclear cells were isolated from recipient spleen or liver as previously described and stained for surface markers and intracellular cytokines using standard flow cytometric protocols [10 (link),11 (link)]. Stained cells were analyzed using FACSDiva software, LSR II (BD Biosciences, San Jose, CA), and FlowJo (Tree Star, Ashland, OR).The following Abs were used for cell-surface staining: anti-CD4–V450, -APC, and -PEcy7 (BD Biosciences), anti-CD8-PEcy5, -APCcy7 and -AF700 (BD Biosciences,); anti–CD45.1-FITC, - and -APC (BD Biosciences). Intracellular staining was carried out using anti-IFN-γ-PE or Per-cp 5.5 (XMG1.2; BD Biosciences), anti–IL-4–PE (11B11; BD Pharmingen), anti–IL-5–PE (TRFK5; BD Pharmingen), anti-Foxp3–PE (FJK-16s; eBioscience).

Bastian D., Liu Y., Wu Y., Schutt S., Nguyen H.D., Daenthanasanmak A., Sofi M.H., Zhang M., Iamsuwat S, & Yu X.Z. (2018). IL-27 Receptor Signaling on T cells Augments GVHD Severity through Enhancing Th1 Responses. Journal of immunology research and therapy, 3(1), 151-157.

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FoxP3 Detection by Flow Cytometry

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For detection of FoxP3, FcR’s were blocked followed by cell surface staining with anti-CD3-PE-Cy7 (145-2C11), anti-CD4-PerCP-Cy5.5 (GK1.5), anti-CD25-APC (PC61.5), anti-IL-10-FITC (JESS-16E3) and anti-FoxP3-PE (FJK-16s) (all from eBioscience).

Bjarnadóttir K., Benkhoucha M., Merkler D., Weber M.S., Payne N.L., Bernard C.C., Molnarfi N, & Lalive P.H. (2016). B cell-derived transforming growth factor-β1 expression limits the induction phase of autoimmune neuroinflammation. Scientific Reports, 6, 34594.

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Profiling Immune Cell Subsets

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Cells isolated from the CNS and spleen of immunized animals were stained extracellularly with anti-CD4-FITC, anti-CD8-V450, anti-CD11b-PerCP, anti-CD11c-PerCP, and anti-CD19-PerCP for 30 minutes on ice. Cells were fixed and permeabilized using Foxp3/Transcription Factor Staining Buffer set (eBioscience) per manufacturer's instructions and then stained intracellularly using anti-Foxp3-PE (FJK-16s, eBioscience) for 30 minutes on ice.

Kersh A.E., Edwards L.J, & Evavold B.D. (2014). Progression of relapsing-remitting demyelinating disease does not require increased TCR affinity or epitope spread. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4429-4438.

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Multiparameter Flow Cytometry Immunophenotyping

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A suspension containing 2 × 10⁶ cells/mL was prepared in PBS and distributed into 96-well round-bottom plates. After centrifugation, the supernatant was discarded and the cells were stained with the viability reagent LIVE/DEAD™ (Thermo Fisher Scientific) and incubated for 10 minutes at 4 °C in the dark. After washes, the cells were incubated for 30 minutes, under the same conditions, with the fluorochrome-conjugated monoclonal antibodies anti-CD3-FITC (clone: 145-2c11), anti-CD19-APC (1D3), anti-CD4-APC-Cy7 (GK1.5), and anti-CD8-PE-Cy5 (53-6.7) (BD Biosciences). After rinsing, the cells were fixed and permeabilized using Foxp3/Transcription Factor Staining Buffer Set (eBioscience, Carlsbad, California, USA) and stained for 1 hour with anti-Foxp3-PE (FJK16s) monoclonal antibody according to the manufacturer’s instructions. The samples were washed and resuspended in PBS 2% FBS, transferred to polypropylene tubes (12 × 75 mm), and acquired by a flow cytometer (LSRFortessa™ X-20, BD Biosciences). Dot plot gating was performed based on cell size (forward scatter) and granularity (side scatter) for lymphocytes, dead cells were excluded (Supplementary Figure 1), and the results were analyzed using FlowJo software (version 7.5.5, BD Biosciences).

Barros R.D., Queiroz L.A., de Assis J.B., Pantoja K.C., Bustia S.X., de Sousa E.S., Rodrigues S.F., Akamine E.H., Sá-Nunes A, & Martins J.O. (2024). Effects of low-dose rapamycin on lymphoid organs of mice prone and resistant to accelerated senescence. Frontiers in Immunology, 15, 1310505.

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Flow Cytometry for Immune Cell Analysis

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Multicolor Flow Cytometry Antibody Panel

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The following fluorophore-conjugated antibodies were purchased from BD Biosciences: anti-CD4 Alx700 (RM4-5), anti-CD4 Alx647 (RM4-5), anti-CD8 PerCP (53–6.7), anti-CD11a (LFA-1) PE-Cy7 (2D7), anti-CD90.1 (Thy1.1) APC-Cy7 (OX-7), anti-CD127 Alx700 (A7R34), anti-CD62L PE (Mel-14), anti-CD44 APC (IM7), anti-IFN-γ APC, and anti-IL-17A PE. Anti-CD25 APC (PC61.5) and anti-Foxp3 PE (FJK-16s) were purchased from eBioscience, Inc. Goat pAb anti-GFP-FITC was purchased from Abcam. Anti-Fcγ-R (2.4G2) was produced from a hybridoma.

Harris M.G., Hulseberg P., Ling C., Karman J., Clarkson B.D., Harding J.S., Zhang M., Sandor A., Christensen K., Nagy A., Sandor M, & Fabry Z. (2014). Immune privilege of the CNS is not the consequence of limited antigen sampling. Scientific Reports, 4, 4422.

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Multiparameter Flow Cytometry of T Cells

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Cells were stained with indicated antibodies in PBS containing 0.2% BSA. For extracellular staining the following fluorescently labeled antibodies were used: Anti-CD4-PerCP (GK1.5, eBioscience, Thermo Fisher Scientific, Waltham, MA, USA), anti-CD44-Pacific Blue (IM7, DRFZ), anti-CD69-FITC (H1.2F3, eBioscience) and anti-CD25-APC (PC61.5, eBioscience). For intracellular staining cells were fixed with fixation buffer and afterwards permeabilized with perm buffer (Foxp3/transcription factor staining buffer set, eBioscience) and then stained with anti-FoxP3-PE (FJK16s, eBioscience), anti-Helios-APC (22F6, Biolegend, San Diego, CA, USA) and anti-Ki67-Pe-Cy7 (B56, BD Bioscience) with the appropriate buffers according to the manufacturer’s protocol. Cells were stored in PBS containing 0.2% BSA and 0.01% sodium azide at 4 °C in the dark until measurement. Flow cytometry was performed using MACS Quant (Miltenyi Biotech, Bergisch Gladbach, Germany) and data were analyzed using FlowJo V 9.6.1 (BD Bioscience).
Gating strategy: 1. Lymphocyte gate in the FSC/SSC plot. 2. Gating either for CD4+FoxP3+ Treg or CD4+FoxP3− Tcon in a CD4/FoxP3 dot plot. 3. Frequencies of CD25+, CD69+, CD44hi, Ki67+ and Helios+ cells among gated CD4+FoxP3+ or CD4+FoxP3− cells were determined either by using different dot plot combinations or by using histograms.

Rose A., von Spee-Mayer C., Kloke L., Wu K., Kühl A., Enghard P., Burmester G.R., Riemekasten G, & Humrich J.Y. (2019). IL-2 Therapy Diminishes Renal Inflammation and the Activity of Kidney-Infiltrating CD4+ T Cells in Murine Lupus Nephritis. Cells, 8(10), 1234.

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