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Trifluorocarbonylcyanide phenylhydrazone fccp

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Trifluorocarbonylcyanide phenylhydrazone (FCCP) is a chemical compound used in laboratory research. It is a protonophore, which means it can transport protons across cell membranes. FCCP is commonly used as a research tool to study mitochondrial function and energy metabolism.

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Lab products found in correlation

Oligomycin, by Agilent Technologies (3 mentions) Antimycin a, by Agilent Technologies (3 mentions) Cyquant cell proliferation assay kit, by Thermo Fisher Scientific (2 mentions) Cell tak, by BD (2 mentions) Cd3 cd28 beads, by Miltenyi Biotec (2 mentions) Rotenone, by Agilent Technologies (2 mentions) Xf base media, by Agilent Technologies (2 mentions) Xfe96 extracellular flux analyzer, by Agilent Technologies (2 mentions) 24 well xf microplate, by Agilent Technologies (1 mentions)

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Phenylhydrazone (3 citations)

3 protocols using trifluorocarbonylcyanide phenylhydrazone fccp

1

Measuring T Cell Metabolism by Seahorse

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OCR was measured using an XFe96 Extracellular Flux Analyzer (Seahorse Bioscience) as previously published 38 . AutoMACS sorted mouse CD4+ and CD8+ T lymphocytes were attached to XFe96 cell culture plates using Cell-Tak (BD Bioscience) in RPMI media with 11 mM glucose. Cells were activated with 1:1 CD3:CD28 beads (Miltenyi BioTech) and vehicle vs 50 µM linoleic acid was added. Twenty-four hours after activation, cells were incubated in serum-free XF Base Media (Seahorse Bioscience) supplemented with 10 mM glucose, 2 mM pyruvate, and 2 µM glutamine, pH 7.4, along with 50 µM linoleic acid if previously present, for 30 minutes at 37°C in a CO2-free cell culture incubator before beginning the assay. Five consecutive measurements, each representing the mean of 8 wells, were obtained at baseline and following sequential addition of 1.25 µM oligomycin, 0.25 µM trifluorocarbonylcyanide phenylhydrazone (FCCP), and 1 µM each of rotenone and antimycin A (all drugs from Seahorse Bioscience). OCR values were normalized to cell number as measured by the CyQUANT Cell Proliferation Assay Kit (Life Technologies).
Ma C., Kesarwala A.H., Eggert T., Medina-Echeverz J., Kleiner D.E., Jin P., Stroncek D.F., Terabe M., Kapoor V., ElGindi M., Han M., Thornton A.M., Zhang H., Egger M., Luo J., Felsher D.W., McVicar D.W., Weber A., Heikenwalder M, & Greten T.F. (2016). NAFLD causes selective CD4+ T lymphocyte loss and promotes hepatocarcinogenesis. Nature, 531(7593), 253-257.
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2

Metabolic Profiling of RASFC Cells

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Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), reflecting oxidative phosphorylation and glycolysis, respectively, were measured before and after treatment with oligomycin (2 μg/mL, Seahorse Biosciences, UK), trifluorocarbonylcyanide phenylhydrazone (FCCP) (5 μM, Seahorse Biosciences) and antimycin A (2 μM, Seahorse Biosciences). RASFC were seeded at 30,000 cells per well in 24-well XF-microplates (Seahorse Biosciences) and allowed to adhere. Cells were then subsequently cultured with Pam3CSK4 (1 μg/ml) for 24 hrs. Following this, cells were rinsed in assay medium (unbuffered Dulbecco’s Modified Eagle’s medium (DMEM)) supplemented with 10 mM glucose, pH7.4) before incubation with assay medium for 30 min at 37 °C in a non-CO2 incubator. Four baseline OCR and ECAR measurements were obtained over 28 min before injection of specific metabolic inhibitors. Moreover, to challenge the metabolic capacity of the RASFC, three OCR and ECAR measurements were obtained over 15 min following injection with oligomycin, FCCP and antimycin A.
McGarry T., Biniecka M., Gao W., Cluxton D., Canavan M., Wade S., Wade S., Gallagher L., Orr C., Veale D.J, & Fearon U. (2017). Resolution of TLR2-induced inflammation through manipulation of metabolic pathways in Rheumatoid Arthritis. Scientific Reports, 7, 43165.
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3

Measuring T Cell Metabolism by Seahorse

Check if the same lab product or an alternative is used in the 5 most similar protocols
OCR was measured using an XFe96 Extracellular Flux Analyzer (Seahorse Bioscience) as previously published 38 . AutoMACS sorted mouse CD4+ and CD8+ T lymphocytes were attached to XFe96 cell culture plates using Cell-Tak (BD Bioscience) in RPMI media with 11 mM glucose. Cells were activated with 1:1 CD3:CD28 beads (Miltenyi BioTech) and vehicle vs 50 µM linoleic acid was added. Twenty-four hours after activation, cells were incubated in serum-free XF Base Media (Seahorse Bioscience) supplemented with 10 mM glucose, 2 mM pyruvate, and 2 µM glutamine, pH 7.4, along with 50 µM linoleic acid if previously present, for 30 minutes at 37°C in a CO2-free cell culture incubator before beginning the assay. Five consecutive measurements, each representing the mean of 8 wells, were obtained at baseline and following sequential addition of 1.25 µM oligomycin, 0.25 µM trifluorocarbonylcyanide phenylhydrazone (FCCP), and 1 µM each of rotenone and antimycin A (all drugs from Seahorse Bioscience). OCR values were normalized to cell number as measured by the CyQUANT Cell Proliferation Assay Kit (Life Technologies).
Ma C., Kesarwala A.H., Eggert T., Medina-Echeverz J., Kleiner D.E., Jin P., Stroncek D.F., Terabe M., Kapoor V., ElGindi M., Han M., Thornton A.M., Zhang H., Egger M., Luo J., Felsher D.W., McVicar D.W., Weber A., Heikenwalder M, & Greten T.F. (2016). NAFLD causes selective CD4+ T lymphocyte loss and promotes hepatocarcinogenesis. Nature, 531(7593), 253-257.
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