In addition to the anti-CSPG4 mAbs (APC labeled), the following fluorochrome-labeled specific for various antigens carried by exosomes were used: Anti-Ox40L-APC (clone 159403, catalog# FAB10541A, conc. 10ug/mL, staining with 10uL/0,1ug, R&D Systems); anti-CD81-APC (clone 1D6, catalog# 17-0819-42, conc. 200ug/mL, staining with 5uL/1ug, Ebioscience,); anti-perforin-PE (clone delta GG9, calatolg# 12-9994-42, conc. 12ug/mL, staining with 5ul/0.06ug, Ebioscience); anti-TGFβ-PE (clone 9016, catalog# FAB2463P, conc. 25ug/mL, staining with 2,5uL/0.06ug, R&D Systems); anti-MIP-1β (Clone D21–1351, catalog# Anti-Ox40L-APC (clone #159403, R & D Systems); anti-CD81 (clone 1D6) APC labeled (Ebioscience); anti-perforin-PE (clone delta GG9), (Ebioscience); anti-TGFβ-PE (clone 9016) (R & D Systems); anti-MIP-1β (clone24006) (R & D Systems.)
Anti perforin pe
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-perforin-PE is a flow cytometry reagent designed to detect the expression of perforin, a cytotoxic protein found in the granules of cytotoxic T cells and natural killer cells. It is conjugated to the fluorescent dye phycoerythrin (PE) for direct detection by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti tgfβ pe, by R&D Systems (1 mentions)
Anti tnf α percp cy5, by Thermo Fisher Scientific (1 mentions)
Anti ifn γ pe cy7, by Thermo Fisher Scientific (1 mentions)
Golgiplug, by BD (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Anti cd3 fitc, by Thermo Fisher Scientific (1 mentions)
Anti ifn γ pe, by Thermo Fisher Scientific (1 mentions)
Anti granzyme b pe, by Thermo Fisher Scientific (1 mentions)
Anti cd56 apc, by Thermo Fisher Scientific (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Pma ionomycin, by BioLegend (1 mentions)
3 protocols using anti perforin pe
1
Exosome Characterization by Flow Cytometry
2
Functional Profiling of HIV-Infected T Cells
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The functional profile response of T cells was evaluated by measuring production of cytokines and effector molecules by flow cytometry. Approximately 1x105 of PBCMs treated with calcitriol or EtOH and infected with HIV-1 from Co-HC, were cultured for 24 hours, in presence of brefeldin (1ug/mL) and monensin (1ug/mL). Extracellular staining for CD4+ and CD8+ T cells was done, as previously described, and intracellular staining was performed using the antibodies anti-IL-2-FITC, anti-IFN-γ-PeCy7, anti-TNF-α-PerCp-Cy5.5, anti-MIP-1β-PE, anti-granzyme-FITC and anti-perforin-PE (eBioscience).
3
Flow Cytometric Analysis of NK Cell Markers
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The cells were stained using the antibodies in Additional file: Table S2 in the dark at 4 °C for 20 min. For intracellular staining, cells were stained with anti-Perforin-PE (eBiosciences, San Diego, CA, USA) or anti-Granzyme B-PE (eBiosciences), and they were fixed and permeabilized using BD CytoFix/CytoPerm™ (BD Biosciences). For evaluation of IFN-γ production, pNK cells were incubated in a 5% CO2 atmosphere with the addition of PMA/Ionomycin (BioLegend, San Diego, CA, USA) and BD GolgiPlug™ (BD Biosciences) at 37 °C for 4 h. Cells were stained with anti-CD3-FITC (eBiosciences) and anti-CD56-APC (eBiosciences), further stained with anti-IFN-γ-PE (eBiosciences), and fixed and permeabilized using BD CytoFix/CytoPerm™(BD Biosciences). Stained cells were analyzed using CytoFLEX flow cytometer (Beckman Coulter, Brea, CA, USA), and data was analyzed using FlowJo version 10.1 software (Treestar Inc., Ashland, OG, USA).
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