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Xf96 polystyrene cell culture microplates

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The XF96 polystyrene cell culture microplates are designed for high-throughput cellular analysis. They provide a 96-well format for culturing and assaying cells. The microplates are made of polystyrene, a common material used in cell culture applications.

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Lab products found in correlation

Xf96 extracellular flux analyzer, by Agilent Technologies (6 mentions) Unbuffered xf assay medium, by Agilent Technologies (3 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Xf assay medium, by Agilent Technologies (2 mentions) Bca assay, by Thermo Fisher Scientific (2 mentions) Bafilomycin a1, by Merck Group (2 mentions) Oligomycin, by Merck Group (2 mentions) Antimycin a, by Merck Group (2 mentions) Chloroquine, by Merck Group (2 mentions) Isoproterenol, by Merck Group (2 mentions)

Close spelling variants of this product

XF96 cell culture microplates (132 citations) XF96 microplates (46 citations) Cell culture microplate (12 citations) Cell-Tak™ coated XF96 polystyrene cell culture microplates (4 citations) XF96 culture microplates (2 citations) Polystyrenes (2 citations)

13 protocols using xf96 polystyrene cell culture microplates

1

Extracellular Flux Analysis of Cultured Cells

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35,000 cells were plated on XF96 polystyrene cell culture microplates (Seahorse®, Agilent, CA, USA) 24 h before the experiment. For the KD experiments, cells were transfected in 10 cm dishes over-night and seeded on XF96 polystyrene cell culture microplates 24 h before the experiment. Thirty minutes before the measurement, cell medium was changed to XF assay medium supplemented with 1 mM sodium pyruvate, 2 mM glutamine and 5.5 mM D-glucose and incubated in non-CO2 37 °C incubator. Prior to loading the plate, XF assay media was refreshed and Ca2+ free assay media was added to respective wells. XF96 extracellular flux analyzer was used to measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). OCR (pmol O2/min) and ECAR (mpH/min) values were normalized to protein content. For the rescue, ECAR was assessed following injection of 25 µl 7x Ca2+ buffer after basal ECAR in 0 Ca2+ was measured. Protein concentration of each well was determined with the PierceTM BCA Protein Assay Kit (Thermo Scientific, Rockford, IL, USA).
Koshenov Z., Oflaz F.E., Hirtl M., Gottschalk B., Rost R., Malli R, & Graier W.F. (2022). Citrin mediated metabolic rewiring in response to altered basal subcellular Ca2+ homeostasis. Communications Biology, 5, 76.
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2

Extracellular Flux Analysis of Cellular Metabolism

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One day before the experiment cells were plated on XF96 polystyrene cell culture microplates (Seahorse®, Agilent, CA, USA). They had to be 100% confluent on the day of the experiment. Before the measurement cells were washed and incubated in XF assay medium supplemented with 1 mM sodium pyruvate, 2 mM glutamine and 5.5 mM D-glucose. An XF96 extracellular flux analyzer was used to measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). OCR (pmol O2/min) and ECAR (mpH/min) values were normalized to protein content.
Depaoli M.R., Karsten F., Madreiter-Sokolowski C.T., Klec C., Gottschalk B., Bischof H., Eroglu E., Waldeck-Weiermair M., Simmen T., Graier W.F, & Malli R. (2018). Real-Time Imaging of Mitochondrial ATP Dynamics Reveals the Metabolic Setting of Single Cells. Cell reports, 25(2), 501-512.e3.
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3

Extracellular Flux Analysis of Cellular Metabolism

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One day before the experiment cells were plated on XF96 polystyrene cell culture microplates (Seahorse®, Agilent, CA, USA). They had to be 100% confluent on the day of the experiment. Before the measurement cells were washed and incubated in XF assay medium supplemented with 1 mM sodium pyruvate, 2 mM glutamine and 5.5 mM D-glucose. An XF96 extracellular flux analyzer was used to measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). OCR (pmol O2/min) and ECAR (mpH/min) values were normalized to protein content.
Depaoli M.R., Karsten F., Madreiter-Sokolowski C.T., Klec C., Gottschalk B., Bischof H., Eroglu E., Waldeck-Weiermair M., Simmen T., Graier W.F, & Malli R. (2018). Real-Time Imaging of Mitochondrial ATP Dynamics Reveals the Metabolic Setting of Single Cells. Cell reports, 25(2), 501-512.e3.
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4

Mitochondrial Respiration in Cell Lines

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INS-1, MIN-6, HeLa and EA.hy926 cells, treated or not with siRNA against presenilin-1 or GSK3β inhibitor CHIR99021, were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience) at a density of 60.000 cells/well for HeLa and EA.hy926, 120.000 cells/well for MIN-6 and 140.000 cells/well for INS-1 cells. After overnight incubation, mitochondrial respiration was performed using an XF96 extracellular flux analyzer (Seahorse Bioscience®) as previously described [21 (link)]. Oxygen consumption was normalized to protein content (pmol O2/min × μg protein).
Klec C., Madreiter-Sokolowski C.T., Stryeck S., Sachdev V., Duta-Mare M., Gottschalk B., Depaoli M.R., Rost R., Hay J., Waldeck-Weiermair M., Kratky D., Madl T., Malli R, & Graier W.F. (2019). Glycogen Synthase Kinase 3 Beta Controls Presenilin-1-Mediated Endoplasmic Reticulum Ca2+ Leak Directed to Mitochondria in Pancreatic Islets and β-Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 52(1), 57-75.
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5

Mitochondrial Respiration in Cell Lines

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INS-1, MIN-6, HeLa and EA.hy926 cells, treated or not with siRNA against presenilin-1 or GSK3β inhibitor CHIR99021, were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience) at a density of 60.000 cells/well for HeLa and EA.hy926, 120.000 cells/well for MIN-6 and 140.000 cells/well for INS-1 cells. After overnight incubation, mitochondrial respiration was performed using an XF96 extracellular flux analyzer (Seahorse Bioscience®) as previously described [21 (link)]. Oxygen consumption was normalized to protein content (pmol O2/min × μg protein).
Klec C., Madreiter-Sokolowski C.T., Stryeck S., Sachdev V., Duta-Mare M., Gottschalk B., Depaoli M.R., Rost R., Hay J., Waldeck-Weiermair M., Kratky D., Madl T., Malli R, & Graier W.F. (2019). Glycogen Synthase Kinase 3 Beta Controls Presenilin-1-Mediated Endoplasmic Reticulum Ca2+ Leak Directed to Mitochondria in Pancreatic Islets and β-Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 52(1), 57-75.
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6

Extracellular Flux Analysis of Cellular Respiration

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Cells were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience) at a density of 30,000 cells per well. After an overnight incubation, cells were washed and preincubated for 30 minutes in unbuffered XF assay medium (Seahorse Bioscience) supplemented with 5.5 mM d-glucose and 1 mM sodium pyruvate at 37°C in a non-CO2 environment. Oxygen consumption rates were subsequently measured using an XF96 extracellular flux analyzer.
Deak A.T., Blass S., Khan M.J., Groschner L.N., Waldeck-Weiermair M., Hallström S., Graier W.F, & Malli R. (2014). IP3-mediated STIM1 oligomerization requires intact mitochondrial Ca2+ uptake. Journal of Cell Science, 127(13), 2944-2955.
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7

Measuring Cell Respiration Rates

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Cells were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience) at a density of 30000 cells per well. After an overnight incubation, cells were washed and preincubated for 30 min in unbuffered XF assay medium (Seahorse Bioscience) supplemented with 5.5 mM D-glucose and 1 mM sodium pyruvate at 37°C in a non-CO2 environment. Oxygen consumption rates were subsequently measured using an XF96 extracellular flux analyzer.
Deak A.T., Blass S., Khan M.J., Groschner L.N., Waldeck-Weiermair M., Hallström S., Graier W.F, & Malli R. (2014). IP3-mediated STIM1 oligomerization requires intact mitochondrial Ca2+ uptake. Journal of Cell Science, 127(13), 2944-2955.
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8

Adipocyte Respiration and Lipid Metabolism

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iBACs were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience®, Agilent, Santa Clara, CA, USA) at a density of 10,000 cells per well and differentiated to mature adipocytes. Pretreatment with inhibitors was performed for 2 h with the following concentrations: 40 μM atglistatin [31 ], 100 nM bafilomycin A1 (Merck, Darmstadt, Germany), and 40 μM chloroquine (Merck, Darmstadt, Germany). All inhibitors and 10 μM isoproterenol (Merck, Darmstadt, Germany) were present during the 30 min equilibration in unbuffered XF assay medium (Seahorse Bioscience®, Agilent, Santa Clara, CA, USA) supplemented with 5.5–25 mM d-glucose, 2 mM glutamine and 1 mM sodium pyruvate at 37 °C in a non-CO2 environment and during measurement. Oxygen consumption rate was subsequently measured every 7 min using an XF96 extracellular flux analyzer (Seahorse Bioscience, Agilent, Santa Clara, CA, USA). Optimal concentrations of specific inhibitors/accelerators of the electron transport chain were determined in a prior titration experiments and used as followed: 2 μM oligomycin, 0.3 μM carbonyl-cyanide p-trifluoromethoxyphenylhydrazone, 2.5 μM antimycin A (all reagents form Merck, Darmstadt, Germany). Results were normalized to protein concentration determined by BCA assay (Thermo Fisher Scientific, Waltham, MA, USA) and presented as pmol O2/(min × μg protein).
Huber K., Hofer D.C., Trefely S., Pelzmann H.J., Madreiter-Sokolowski C., Duta-Mare M., Schlager S., Trausinger G., Stryeck S., Graier W.F., Kolb D., Magnes C., Snyder N.W., Prokesch A., Kratky D., Madl T., Wellen K.E, & Bogner-Strauss J.G. (2018). N-acetylaspartate pathway is nutrient responsive and coordinates lipid and energy metabolism in brown adipocytes. Biochimica et biophysica acta. Molecular cell research, 1866(3), 337-348.
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9

Macrophage Mitochondrial Respiration Assay

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Macrophages were plated in XF96 polystyrene cell culture microplates (Seahorse Bioscience®, North Billerica, MA) at a density of 60,000 cells per well. After 24 h, cells were washed and preincubated for 30 min in XF assay medium supplemented with sodium pyruvate (1 mM) with or without glutamine (2 mM) and glucose (25 mM) at 37°C in a nonCO2 environment. The oxygen consumption rate (OCR) was subsequently measured every 7 min using an XF96 extracellular flux analyzer (Seahorse Bioscience®). A standard protocol with 15 min basal measurement followed by 10 μM oligomycin, addition of 0.3 μM carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), and 2.5 μM antimycin A was performed. Oxygen consumption was either normalized to protein content (pmol O2/min × μg protein) or expressed as a percentage of the maximal mitochondrial respiration in the presence of 0.3 μM FCCP.
Goeritzer M., Schlager S., Radovic B., Madreiter C.T., Rainer S., Thomas G., Lord C.C., Sacks J., Brown A.L., Vujic N., Obrowsky S., Sachdev V., Kolb D., Chandak P.G., Graier W.F., Sattler W., Brown J.M, & Kratky D. (2014). Deletion of CGI-58 or adipose triglyceride lipase differently affects macrophage function and atherosclerosis. Journal of Lipid Research, 55(12), 2562-2575.
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10

Measuring Mitochondrial Respiration in C2C12 Cells

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C2C12 cells were seeded in XF96 polystyrene cell culture microplates (Seahorse Bioscience) at a density of 25.000 cells per well. 24 h after plating, cells were washed and preincubated for 30 min in serum-free XF assay medium supplemented with D-glucose (25 mM), sodium pyruvate (1 mM) and glutamine (2 mM) at 37°C in a non-CO2 environment. Oxygen consumption rate (OCR) was subsequently measured every 7 min using an XF96 extracellular flux analyzer (Seahorse Bioscience). 2.5 μM antimycin A was used to stop mitochondrial respiration (= non-mitochondrial respiration). Basal mitochondrial oxygen consumption rate (OCR) was calculated by subtracting average of 3 subsequent non-mitochondrial OCRs from average of three basal OCRs (prior to antimycin treatment) and normalized to individual protein amount (pmol O2/(min x protein per well)).
For CB-839 treatment, 60,000 C2C12 cells were seeded in XF24-well plates and allowed to adhere overnight. The next day, wells were washed twice with PBS and serum-free, pyruvate-free DMEM, in the presence or absence of 1μM CB-839 (or 0.01% DMSO). 6 hours after treatment, OCRs were measured as described above and subsequently normalized to cell counts and OCRs of individual genotype (control versus AGC1-KD) in DMSO treated conditions.
Alkan H.F., Walter K.E., Luengo A., Madreiter-Sokolowski C.T., Stryeck S., Lau A.N., Al-Zoughbi W., Lewis C.A., Thomas C.J., Hoefler G., Graier W.F., Madl T., Vander Heiden M.G, & Bogner-Strauss J.G. (2018). Cytosolic Aspartate Availability Determines Cell Survival When Glutamine Is Limiting. Cell metabolism, 28(5), 706-720.e6.
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