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Foxp3 apc pch101

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The Foxp3-APC (PCH101) is a fluorescently-labeled antibody that can be used to detect and quantify the expression of the Foxp3 transcription factor in cells. Foxp3 is a key marker for regulatory T cells (Tregs) and is essential for their development and function. The antibody is conjugated to the fluorescent dye Allophycocyanin (APC), which allows for the detection of Foxp3-positive cells by flow cytometry.

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Lab products found in correlation

Cd127 pe, by BD (2 mentions) Cd3 alexa fluor 700, by BD (2 mentions) Cd3 percp cy5, by BD (2 mentions) Cd4 v450, by BD (2 mentions) Cd4 pe, by BD (2 mentions) Cd4 fitc, by BD (2 mentions) Cd4 v500, by BD (2 mentions) Cd25 bv605, by BD (2 mentions) Cd25 pe, by BD (2 mentions) Cd127 percp cy5, by BD (2 mentions) Cd196 bv605, by BD (2 mentions) Il 17a alexa fluor 700, by BD (2 mentions) Foxp3 v450, by BD (2 mentions) Anti human foxp3 staining set, by Thermo Fisher Scientific (2 mentions) Facs lysis, by BD (2 mentions) Brefeldin a, by Merck Group (2 mentions) Rpmi 1640, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Paraformaldehyde (pfa), by Merck Group (2 mentions) Ionomycin, by Merck Group (2 mentions)

5 protocols using foxp3 apc pch101

1

Multicolor Flow Cytometry Analysis

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For multicolor flow cytometric analyses, the following fluorochrome-labeled Ab and IgG subset controls (all purchased from Becton-Dickinson (BD, Franklin Lakes, NJ, unless otherwise indicated) were utilized for surface or intracellular staining of cells: CD3-Alexa Fluor 700 (clone UCHT1), CD3-PerCPCy5.5 (5k7), CD4-V450 (RPA-T4), CD4-PE (RPA-T4), CD4-FITC (RPA-T4), CD4-V500 (RPA-T4), CD25-BV605 (2A3), CD25-PE (2A3), CD127-PE (hIL-7R-M21), CD127-PErCPCy5.5 (hIL-7R-M21), CD196-BV605 (11A9), IL-17A-Alexa Fluor 700 (N49-653), RORγt-PE (Q21-555), FoxP3-V450 (259D/C7). Foxp3-APC (PCH101) was a component of the Anti-Human Foxp3 Staining Set (eBioscience, San Diego, CA). FacsLysis was purchased from BD. The following reagents were purchased from Sigma-Aldrich, Inc. (St. Louis, MO): RPMI-1640, phosphate-buffered saline (PBS), fetal bovine serum, paraformaldehyde, phorbol 12-myristate 13-acetate, ionomycin, brefeldin A.
Rito D.C., Viehl L.T., Buchanan P.M., Haridas S, & Koenig J.M. (2016). Augmented Th17-Type Immune Responses in Preterm Neonates Exposed to Histologic Chorioamnionitis: Chorioamnionitis and Th17-type immunity. Pediatric research, 81(4), 639-645.
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2

Multiparametric Flow Cytometry Analysis

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BALF cells and PBMCs were stained for extra- and intracellular markers using the following antibodies: CD3-APC-eFluor780(SK7), CD4-AF700(OKT4), CD45RO-FITC(UCHl-1), CD95-APC(DX2), FoxP3-APC(PCH101) (eBiosciences) and CD25-PE(M-A251), CD25-PE-Cy7(M-A251), CD127-V450(hIL7R-M21) anti-CTLA-4-BV421(BNI3) (BD biosciences) and CD45RA-PE-Texas Red(MEM-56) (Invitrogen). Fixable Aqua Dead Cell Stain kit for 405 nm (Invitrogen, Molecular Probes) was used as live-dead marker. Cells were measured on a Flow cytometer LSRII (BD Biosciences).
Broos C.E., van Nimwegen M., Kleinjan A., ten Berge B., Muskens F., in ’t Veen J.C., Annema J.T., Lambrecht B.N., Hoogsteden H.C., Hendriks R.W., Kool M, & van den Blink B. (2015). Impaired survival of regulatory T cells in pulmonary sarcoidosis. Respiratory Research, 16(1), 108.
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3

Multicolor Flow Cytometry Analysis

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For multicolor flow cytometric analyses, the following fluorochrome-labeled Ab and IgG subset controls (all purchased from Becton-Dickinson (BD, Franklin Lakes, NJ, unless otherwise indicated) were utilized for surface or intracellular staining of cells: CD3-Alexa Fluor 700 (clone UCHT1), CD3-PerCPCy5.5 (5k7), CD4-V450 (RPA-T4), CD4-PE (RPA-T4), CD4-FITC (RPA-T4), CD4-V500 (RPA-T4), CD25-BV605 (2A3), CD25-PE (2A3), CD127-PE (hIL-7R-M21), CD127-PErCPCy5.5 (hIL-7R-M21), CD196-BV605 (11A9), IL-17A-Alexa Fluor 700 (N49-653), RORγt-PE (Q21-555), FoxP3-V450 (259D/C7). Foxp3-APC (PCH101) was a component of the Anti-Human Foxp3 Staining Set (eBioscience, San Diego, CA). FacsLysis was purchased from BD. The following reagents were purchased from Sigma-Aldrich, Inc. (St. Louis, MO): RPMI-1640, phosphate-buffered saline (PBS), fetal bovine serum, paraformaldehyde, phorbol 12-myristate 13-acetate, ionomycin, brefeldin A.
Rito D.C., Viehl L.T., Buchanan P.M., Haridas S, & Koenig J.M. (2016). Augmented Th17-Type Immune Responses in Preterm Neonates Exposed to Histologic Chorioamnionitis: Chorioamnionitis and Th17-type immunity. Pediatric research, 81(4), 639-645.
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4

Flow Cytometry Analysis of T Cell Subsets

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Whole blood was stained for flow cytometry with multiple combinations of the following mAbs: CD3-FITC (SP34) ; CD20-PE (3G8); CD8-PerCP (SK1); CD4-APC (L200); CD25-FITC (2A3); HLA-DR-PerCP (l243); Ki-67-FITC (B56) (all from BD Biosciences) and FoxP3-APC (PCH101) (EBiosciences), as described (17 (link), 18 (link)). Data were acquired with a FACSCalibur flow cytometer (BD Immunocytometry Systems) and analyzed with CellQuest software (BD Biosciences). CD4+ and CD8+ T cell percentages were obtained by first gating on lymphocytes and then on CD3+ T cells. Immune activation, and proliferation markers were determined by gating on lymphocytes, then on CD3+ T cells, and finally on CD4+CD3+ or CD8+CD3+ T cells.
He T., Brocca-Cofano E., Policicchio B.B., Sivanandham R., Gautam R., Raehtz K.D., Xu C., Pandrea I, & Apetrei C. (2016). T Regulatory Cell Depletion Reactivates Latent SIV in Controller Macaques while Boosting SIV-specific T Lymphocytes. Journal of immunology (Baltimore, Md. : 1950), 197(12), 4535-4539.
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5

Comprehensive Immunophenotyping of Cell Subsets

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The following monoclonal antibodies were purchased from BD Biosciences: 7-AAD, CCR7, CD3, CD4, CD16, CD14, CD19, CD25, CD34, CD45, CD56, CD69, CD94, CD117, CTLA4 and DAPI; from eBiosciences: CD62L and CD127; from R&D Systems: CCR5, CCR6, CXCR1, CXCR3, GITR, integrin β7 and LAP. Titrated amounts of 7AAD and DAPI were added to stained cells 10 minutes before acquisition. For Foxp3 intranuclear staining, cells were then fixed and permeabilized at room temperature for 15 minutes using the Foxp3 Fix/Perm Kit (eBioscience) and stained in permeabilization buffer with a directly labeled antibody to Foxp3 APC (PCH101; eBioscience) at room temperature for 45 minutes, prior to two washes with permeabilization buffer. The LSRFortessa and FACSCalibur (Becton Dickinson) cell analyzers were used to acquire data. FlowJo software (Tree Star) was used for data analysis.
Pedroza-Pacheco I., Shah D., Domogala A., Luevano M., Blundell M., Jackson N., Thrasher A., Madrigal A, & Saudemont A. (2016). Regulatory T cells inhibit CD34+ cell differentiation into NK cells by blocking their proliferation. Scientific Reports, 6, 22097.
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