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Seahorse xf96 polystyrene tissue culture plates

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The Seahorse XF96 polystyrene tissue culture plates are designed for use with Agilent's Seahorse XF Analyzers. They provide a standardized, single-use platform for conducting real-time measurements of cellular metabolic activity.

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Lab products found in correlation

Xf96 extracellular flux analyzer, by Agilent Technologies (3 mentions) Seahorse bioscience xf 96 extracellular flux analyzer, by Agilent Technologies (2 mentions) Xf base medium, by Agilent Technologies (2 mentions) Xfe96 extracellular flux analyzer, by Agilent Technologies (1 mentions) Ensight, by PerkinElmer (1 mentions)

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Seahorse XF96 (83 citations) XF96 plates (49 citations) Seahorse plates (20 citations) Seahorse XF96 plates (16 citations) Seahorse culture plate (5 citations) XF96 Culture Plates (2 citations) Polystyrenes (2 citations)

10 protocols using seahorse xf96 polystyrene tissue culture plates

1

Measuring Cellular Respiration Dynamics

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The rates of oxygen consumption (OCR) in various cell lines were determined with a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience), as detailed elsewhere (46 (link)). Cells were seeded at a density of 1 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience). Inhibitors were used at the following concentrations: Oligomycin (to inhibit the ATP synthase) (1.5 μM), Carbonyl cyanide 4-trifluoromethoxy-phenylhydrazone (FCCP) (to uncouple the mitochondrial inner membrane and allow for maximum electron flux through the electron transfer chain) (0.15μM), Antimycin A (to inhibit complex III) (5 μM) and Rotenone (to inhibit complex I) (1 μM).
Zhao X., Cui L., Xiao Y., Mao Q., Aishanjiang M., Kong W., Liu Y., Chen H., Hong F., Jia Z., Wang M., Jiang P, & Guan M.X. (2019). Hypertension-associated mitochondrial DNA 4401A>G mutation caused the aberrant processing of tRNAMet, all 8 tRNAs and ND6 mRNA in the light-strand transcript. Nucleic Acids Research, 47(19), 10340-10356.
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2

Measuring Mitochondrial Function in Cybrid Cells

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The OCR in various cybrid cell lines were measured with a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience), as detailed previously (50 (link), 53 (link)). Cells were seeded at a density of 2 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience). Inhibitors were used at the following concentrations: oligomycin (1.5 μM), FCCP (0.8 μM), antimycin A (1.5 μM), and rotenone (3 μM), respectively.
Ji Y., Nie Z., Meng F., Hu C., Chen H., Jin L., Chen M., Zhang M., Zhang J., Liang M., Wang M, & Guan M.X. (2021). Mechanistic insights into mitochondrial tRNAAla 3’-end metabolism deficiency. The Journal of Biological Chemistry, 297(1), 100816.
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3

Extracellular Acidification Rate Analysis

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Real-time changes in extracellular acidification rate (ECAR) of HPAECs were analyzed with an XF96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA, USA) as described previously (27 (link)). Briefly, HPAECs were seeded at 2.5 × 104 per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience) and incubated at 37 °C overnight in VCBM. The next day, after 1 μg/ml LPS treatment for 4 hours, the medium was changed to XF base Medium (Seahorse Bioscience) supplemented with 2 mM glutamine, and then the plate was incubated for 1 h in a non-CO2 incubator at 37 °C. The ECAR assay was performed on the XF96 Extracellular Flux Analyzer (Seahorse Bioscience), and the ECAR values were normalized using protein concentration. Inhibitors and activators were used in these tests at the following concentrations: glucose (10 mM), oligomycin (2 μM), 2-DG (50 mM).
Wang L., Cao Y., Gorshkov B., Zhou Y., Yang Q., Xu J., Ma Q., Zhang X., Wang J., Mao X., Zeng X., Su Y., Verin A., Hong M., Liu Z, & Huo Y. (2019). Ablation of endothelial Pfkfb3 protects mice from acute lung injury in LPS-induced endotoxemia. Pharmacological research, 146, 104292.
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4

Measuring Cellular Oxygen Consumption

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The oxygen consumption rate (OCR) in various cell lines were measured with a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience), as detailed previously [42 (link)]. Cells from each cell line were seeded at a density of 2 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience, North Billerica, Massachusetts). Inhibitors for various OXPHOS complexes were used at the following concentrations: 1 μM of oligomtcin (to inhibit the ATP synthase), 0.5  μM of carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) (to uncouple the mitochondrial inner membrane and allow for maximum electron flux through the ETC), 1 μM of rotenone (to inhibit complex I), and 1 μM of antimycin A (to inhibit complex III), respectively.
Wang J., Ji Y., Ai C., Chen J.R., Gan D., Zhang J., Mo J.Q, & Guan M.X. (2023). Optimized allotopic expression of mitochondrial ND6 transgene restored complex I and apoptosis deficiencies caused by LHON-linked ND6 14484T > C mutation. Journal of Biomedical Science, 30, 63.
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5

Monocyte Metabolic Profiling Using Seahorse

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FACS-sorted Ly6Chi and Ly6Clo monocytes from spleens were seeded in quintuplicate in Seahorse XF96 polystyrene tissue culture plates (2 × 105 monocytes per well) (Seahorse Bioscience). The metabolic rates of monocytes were analysed in four consecutive measurements in XF Base Medium (unbuffered DMEM with 5.5 mM glucose and 2 mM L-glutamine, pH adjusted to 7.4). After three basal measurements, three consecutive measurements were taken following the addition of 10 mM glucose, 2 μM oligomycin, and 50 mM 2-DG to determine basal and maximum ECAR. All compounds used during the Seahorse runs were from Sigma-Aldrich.
Karle I.L., Perozzo M.A., Mishra V.K, & Balaram P. (1998). Crystal structure of the channel-forming polypeptide antiamoebin in a membrane-mimetic environment. Proceedings of the National Academy of Sciences of the United States of America, 95(10), 5501-5504.
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6

Measuring Cellular Oxygen Consumption

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The rates of oxygen consumption (OCR) in various cell lines were measured with a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience), as detailed previously (44 (link),48 (link)). Cybrid cells were seeded at a density of 2 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience). Inhibitors were used at the following concentrations: Oligomycin (1.5 μM), Carbonyl cyanide 4-trifluoromethoxy-phenylhydrazone (FCCP) (0.8 μM), Antimycin A (1.5 μM) and Rotenone (3 μM).
Jia Z., Zhang Y., Li Q., Ye Z., Liu Y., Fu C., Cang X., Wang M, & Guan M.X. (2018). A coronary artery disease-associated tRNAThr mutation altered mitochondrial function, apoptosis and angiogenesis. Nucleic Acids Research, 47(4), 2056-2074.
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7

Measuring Oxygen Consumption in Cybrid Cells

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The rates of oxygen consumption (OCR) in various cybrid cell lines were measured with a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience), as detailed previously (61 (link),62 (link)). Cybrid cells were seeded at a density of 2 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience). Inhibitors were used at the following concentrations: oligomycin (1.5 μM), carbonyl cyanide 4-trifluoromethoxy-phenylhydrazone (FCCP) (0.8 μM), antimycin A (1.5 μM) and rotenone (3 μM).
Meng F., Zhou M., Xiao Y., Mao X., Zheng J., Lin J., Lin T., Ye Z., Cang X., Fu Y., Wang M, & Guan M.X. (2021). A deafness-associated tRNA mutation caused pleiotropic effects on the m1G37 modification, processing, stability and aminoacylation of tRNAIle and mitochondrial translation. Nucleic Acids Research, 49(2), 1075-1093.
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8

Extracellular Flux Analysis of HRMECs

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HRMECs were seeded on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience, North Billerica, MA), and incubated at 37 °C overnight in 25% VCBM. To avoid differences due to unequal cell numbers and growth rates, all measurements were made starting with confluent cells by seeding 1.5 × 104 per well. The next day, the medium was changed to XF base Medium (Seahorse Bioscience) supplemented with 2 mM glutamine (for ECAR), or supplemented with 25 mM glucose, 1 mM pyruvate, and 2 mM glutamine (for oxygen consumption rate, OCR), and then the plate was incubated for 1 h in a non-CO2 incubator at 37 °C. ECAR and OCR were measured with an XFe96 extracellular flux analyzer (Seahorse Bioscience). Inhibitors and activators were used in these tests at the following concentrations: glucose (10 mM), oligomycin (2 µM), 2-DG (50 mM), FCCP (1 µM), antimycin A (0.5µM), and rotenone (0.5 µM).
Liu Z., Yan S., Wang J., Xu Y., Wang Y., Zhang S., Xu X., Yang Q., Zeng X., Zhou Y., Gu X., Lu S., Fu Z., Fulton D.J., Weintraub N.L., Caldwell R.B., Zhang W., Wu C., Liu X.L., Chen J.F., Ahmad A., Kaddour-Djebbar I., Al-Shabrawey M., Li Q., Jiang X., Sun Y., Sodhi A., Smith L., Hong M, & Huo Y. (2017). Endothelial adenosine A2a receptor-mediated glycolysis is essential for pathological retinal angiogenesis. Nature Communications, 8, 584.
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9

Seahorse Analyzer for OCR and ECAR

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OCR and ECAR measurements were performed using a Seahorse Bioscience XF-96 extracellular flux analyzer (Seahorse Bioscience). Cells at a density of 2 × 104 per well were seeded onto Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience) until they adhered. The cells were washed twice with 1 ml of pre-warmed assay medium (DMEM medium plus 1.85 g/l NaCl, 10 mM glucose, 1 mM pyruvate, and 2 mM glutamine, pH 7.4). After washing, 500 μl of Assay Medium was added per well, and the 24-well plate was placed in a CO2-free 37°C incubator for 30 min. Subsequently, 75 μl of oligomycin, FCCP, rotenone and antimycin A (final concentration of 1 μM) were added to the four channels per well according to the manufacturer's instructions. The ECAR assay procedure was similar to that used for OCR. However, no glucose or pyruvate was detected in the ECAR assay medium. The following inhibitors were used: 10 mM glucose, 1 μM oligomycin, and 1 μM 2-DG (2-deoxy-d-glucose). The results were normalized by cell counting using EnSight (Perkin Elmer).
Zhang Y., Zhou J.B., Yin Y., Wang E.D, & Zhou X.L. (2024). Multifaceted roles of t6A biogenesis in efficiency and fidelity of mitochondrial gene expression. Nucleic Acids Research, 52(6), 3213-3233.
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10

Measuring Mitochondrial Function in MSCs

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MSCs were plated in a density of 2 × 104 cells per well on Seahorse XF96 polystyrene tissue culture plates (Seahorse Bioscience, North Billerica, MA, USA). Then, Oligomycin (2 μM), Carbonyl cyanide 4-trifluoromethoxy-phenylhydrazone (FCCP) (0.25 μM), Antimycin A (0.5 μM) and Rotenone (3 μM) were delivered to detect the spare respiration capacity, maximal respiration and ATP productivity, respectively.
Wang H., Sun Y., Pi C., Yu X., Gao X., Zhang C., Sun H., Zhang H., Shi Y, & He X. (2022). Nicotinamide Mononucleotide Supplementation Improves Mitochondrial Dysfunction and Rescues Cellular Senescence by NAD+/Sirt3 Pathway in Mesenchymal Stem Cells. International Journal of Molecular Sciences, 23(23), 14739.
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