Seahorse xf culture plates

Manufactured by Agilent Technologies

The Seahorse XF culture plates are specialized multi-well plates designed for use with Agilent's Seahorse XF analyzers. These plates are engineered to provide a controlled and consistent environment for the cultivation and analysis of cells, tissues, or other biological samples during Seahorse XF experiments.

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Lab products found in correlation

Xf24 extracellular flux analyzer, by Agilent Technologies (4 mentions) Xf base medium, by Agilent Technologies (2 mentions) Cyquant kit, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

Seahorse plates (20 citations) Seahorse XF cell culture plates (8 citations) Seahorse culture plate (5 citations)

4 protocols using seahorse xf culture plates

Mitochondrial Function Assay in Neurons

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Mitochondrial function was assessed using the Seahorse Bioscience XF24 Extracellular Flux Analyzer. Neurons were plated on Seahorse XF culture plates (Seahorse Bioscience) at a density of 65,000 cells/well. On day 7, cells were rinsed in assay medium (pH 7.4) containing XF Base medium (Seahorse Bioscience), 5.5 mM glucose, and 1 mM sodium-pyruvate. Cells remained in assay medium 1 h at 37°C in a non-CO2 incubator prior to initializing the Seahorse24XF analysis. Using the MitoStress Kit as previously described [30 (link)], oxygen consumption rate (OCR) was measured under varying conditions. After three initial baseline measurements of OCR, the ATP synthase inhibitor oligomycin (1 μm) was added and three subsequent measurements were taken. Next, an electron transport chain (ETC) accelerator, p-trifluoromethoxy carbonyl cyanide phenyl hydrazone (FCCP at 1.5 μm), was added, and after 3 measurements were taken, mitochondrial inhibitors rotenone (1 μm) and antimycin (1 μm) were added, and three final measurements were taken. Data was normalized to total DNA content, which was determined from each well using the CyQuant kit (Invitrogen) as per the manufacturer's instructions.

Gray N.E., Zweig J.A., Matthews D.G., Caruso M., Quinn J.F, & Soumyanath A. (2017). Centella asiatica Attenuates Mitochondrial Dysfunction and Oxidative Stress in Aβ-Exposed Hippocampal Neurons. Oxidative Medicine and Cellular Longevity, 2017, 7023091.

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Mitochondrial Function Assessment via Seahorse

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Mitochondrial function was assessed using the Seahorse Bioscience XF24 Extracellular Flux Analyzer. Isolated mitochondria were plated on Seahorse XF culture plates (Seahorse Bioscience) at a concentration of 2ug/well with 3-4 replicate wells per animal. The plate was centrifuged for 15 minutes at 2000xg and oxygen consumption rates (OCR) were measured under varying conditions using the MitoStress Kit as previously described (Iuso 2017 (link)). After three initial baseline measurements of OCR, a saturating concentration of ADP (2mM) was added to ensure maximum state III respiration and three subsequent measurements were taken. This was followed by the addition of the ATP synthase inhibitor oligomycin (2 μM) to induce state IV respiration and three additional measurements were taken. Next an electron transport chain (ETC) accelerator carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP at 4 μM), was added to induce maximal uncoupled (state IIIu) respiration and after 3 measurements were taken, mitochondrial inhibitors rotenone (1 μM) and antimycin (1 μM) were added, and three final measurements were taken.

Gray N.E., Zweig J.A., Caruso M., Zhu J.Y., Wright K.M., Quinn J.F, & Soumyanath A. (2018). Centella asiatica attenuates hippocampal mitochondrial dysfunction and improves memory and executive function in β-amyloid overexpressing mice. Molecular and cellular neurosciences, 93, 1-9.

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Evaluating Mitochondrial Function via Seahorse

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Mitochondrial function was evaluated using the Seahorse Bioscience XF24 Extracellular Flux Analyzer. Isolated hippocampal mitochondria were first plated on Seahorse XF culture plates (Seahorse Bioscience) (2 μg/well) with 3–4 replicate wells per animal. The plate was then centrifuged for 15 minutes at 2000g and oxygen consumption rates (OCRs) were quantified under different conditions using the MitoStress Kit as previously described (Iuso et al., 2017 (link)). Following 3 baseline measurements of OCR (reflecting basal respiration), a saturating concentration of ADP (2 mM) was added to achieve maximum state III respiration and 3 successive measurements were taken (denoted as ADP-stimulated respiration). The ATP synthase inhibitor oligomycin (2 μm) was then added to induce state IV respiration and 3 more measurements were made. Next the electron transport chain (ETC) accelerator carbonyl cyanide p-(trifluoromethoxy)phenyl-hydrazone (FCCP at 4 μm) was added to induce maximal uncoupled (state IIIu) respiration and after 3 measurements, the mitochondrial inhibitors rotenone (1 μm) and antimycin (1 μm) were added, and 3 final measurements were made. The respiratory control ratio (RCR) between state III and state IV respiration was also calculated.

Zweig J.A., Brandes M.S., Brumbach B.H., Caruso M., Wright K.M., Quinn J.F., Soumyanath A, & Gray N.E. (2020). Loss of NRF2 accelerates cognitive decline, exacerbates mitochondrial dysfunction, and is required for the cognitive enhancing effects of Centella asiatica during aging. Neurobiology of aging, 100, 48-58.

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Mitochondrial Function Assessment in Alzheimer's

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Mitochondrial function was assessed using the Seahorse Bioscience XF24 Extracellular Flux Analyzer. SH-SY5Y cells were plated on Seahorse XF culture plates (Seahorse Bioscience) in DMEM/F12 containing N2 growth supplement and either CAW (100ug/mL), 1,5dCQA(1.5uM) or IsoA (1.5uM), at 60,000 cells/well, which was determined to be the optimum density for basal O₂ consumption rate (OCR). The following day Aβ_25–35 (50uM) was added. Two days later cells were rinsed in assay medium (pH 7.4) containing XF Base medium (Seahorse Bioscience), 5.5mM glucose and 1mM sodium-pyruvate. Cells remained in assay medium 1h at 37 C in a non-CO2 incubator prior to initializing the Seahorse24XF analysis. Using the MitoStress Kit as previously described [29 (link)], OCR was measured under varying conditions. After three initial baseline measurements of OCR, the ATP synthase inhibitor oligomycin (1 μM) was added and three subsequent measurements were taken. Next an ETC accelerator, p-trifluoromethoxy carbonyl cyanide phenyl hydrazone (FCCP at 1.5 μM), was added and after 3 measurements were taken, mitochondrial inhibitors rotenone (1 μM) and antimycin (1 μM) were added, and three final measurements were taken. Data was normalized to total DNA content, which was determined from each well using the CyQuant kit (Invitrogen) as per the manufacturer’s instructions.

Gray N.E., Sampath H., Zweig J.A., Quinn J.F, & Soumyanath A. (2015). Centella asiatica attenuates β-amyloid-induced oxidative stress and mitochondrial dysfunction. Journal of Alzheimer's disease : JAD, 45(3), 933-946.

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