Fccp for seahorse
WebThis covers instrument time, the Seahorse cartridge plate, the Seahorse microplate for cell seeding, Seahorse media, reagents for a standard Mitochondrial Stress Test (MST) or … WebAug 6, 2024 · Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) is an uncoupling agent that collapses the proton gradient and disrupts the mitochondrial membrane potential. ... Although the Seahorse ...
Fccp for seahorse
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WebIn this chapter, we outline a detailed protocol to measure the rate of oxygen consumption (OCR) with the high-throughput analyzer Seahorse XF96. More importantly, this protocol … WebDilution volumes for FCCP - Cell Characterization with XFp Ports Used for FCCP Volume of Assay Media (µl) Volume of Stock FCCP (µl) 10X Final FCCP (Port) Concentra-tion (µM) Low Range B, C 487.5 12.5 1.25 D 475 25 2.5 High Range B, C 450 50 5 D 400 100 10 Alternatively, serial dilutions of FCCP may be prepared by add-
WebThe XF Cell Energy Phenotype Test was performed by injecting oligomycin + FCCP (1.0 μM and 0.5 μM final, respectively) and including 20 μM Hoechst 33342 (2.0 μM final). A Turnkey Solution That Integrates Cell Count Normalization Into Wave Software WebYes, Seahorse provides a very narrow and restricted snapshot of mitochondrial function. When using the Seahorse protocol subsequent injections of ADP, Oligomycin, and …
WebJan 7, 2024 · FCCP, as an uncoupling agent of oxidative phosphorylation, could reverse the hyperpolarized state caused by oligomycin through carrying the protons across the mitochondrial inner membrane. Finally, the injection of R/A could inhibit mitochondrial complexes I and III completely stopping the mitochondrial respiration. WebThe XF Cell Energy Phenotype Test was performed by injecting oligomycin + FCCP (1.0 μM and 0.5 μM final, respectively) and including 20 μM Hoechst 33342 (2.0 μM final). A Turnkey Solution That Integrates Cell …
WebCryptococcus neoformans is a human fungal pathogen that adapts its metabolism to cope with limited oxygen availability, nutrient deprivation and host phagocytes. To gain insight into cryptococcal metabolism, we optimized a protocol for the Seahorse Analyzer, which measures extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) as …
WebDec 9, 2024 · Real-time measurement of tumor cell glycolytic rate using a Seahorse extracellular flux analyzer showed a much lower extracellular acidification rate (ECAR) of KO PyVT cells compared to WT PyVT cells ... 0.5 μM FCCP, and 1 μM antimycin/rotenone. Western blot and quantitative real-time PCR (qRT-PCR) analyses ... bebe gateando para dibujarWebOct 15, 2024 · Subsequently, cells were equilibrated in the Seahorse analyzer for 20 min at 37 °C and baseline OCR was measured, after which 1.6 µM oligomycin A (port A), 1.1 µM FCCP (port B), and 4.62 μM antimycin A/2.31 μM rotenone (port C) were sequentially added (all chemicals from Sigma-Aldrich). bebe gateandoWebNext carbonyl cyanide-p-trifluoromethox- yphenyl-hydrazon (FCCP), a protonophore, is added to collapse the inner membrane gradient, allowing the ETC to function at its … displej nereaguje na dotyk samsungWebHas anyone seen no response of HEK293 cells to FCCP in a Seahorse assay? Hello, I performed a titration of FCCP in HEK cells and two other melanoma cells lines in order to … displej mobilu nereagujeWebSeahorse Bioscience Extracellular Flux Analyzer. The Seahorse XF-96 measures the rate of change of analytes dissolved oxygen and pH in media immediately surrounding living cells cultured in a microplate. Changes in the extracellular media are caused by the consumption or production of analytes by the cells. Therefore, a sensitive measurement … bebe gatoWebIn the lab we tried to adapt the SeaHorse mitochondrial stress protocol using bulk metabolic inhibitors from Sigma. At the beginning everything worked totally fine, but later on we found out that... bebe gateando facil de dibujarWebApr 7, 2024 · The hypoxia associated with the transforming growth factor-β2 (TGF-β2)-induced epithelial mesenchymal transition (EMT) of human retinal pigment epithelium (HRPE) cells is well recognized as the essential underlying mechanism responsible for the development of proliferative retinal diseases. In vitro, three … displej za iphone 7 plus