Log In Sign up
The largest database of trusted experimental protocols

Extracellular flux analysis

Manufactured by Agilent Technologies
Visit Supplier
Sourced in United States

The Extracellular Flux Analysis is a lab equipment product that measures the metabolic activity of cells. It quantifies the oxygen consumption rate and extracellular acidification rate of cells in real-time.

Automatically generated - may contain errors

Lab products found in correlation

Xf24 well microplate, by Agilent Technologies (3 mentions) Oligomycin, by Merck Group (3 mentions) Glucose, by Thermo Fisher Scientific (2 mentions) Cell tak, by Corning (2 mentions) L glutamine, by Merck Group (2 mentions) Antimycin a, by Merck Group (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Rotenone, by Merck Group (1 mentions) Rotenone antimycin a, by Agilent Technologies (1 mentions)

7 protocols using extracellular flux analysis

1

Extracellular Flux Analysis of Cellular Metabolism

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Real-time changes in extracellular acidification rates (ECARs, as a measure of lactate production) and oxygen consumption rates (OCRs, as a measure of oxidative phosphorylation) were analyzed using extracellular flux analysis (Seahorse Bioscience, North Billerica, MA) as described previously [48] (link). Briefly, UT and CE J774 cells were plated at 1.5x105 cells per well in 50 µl of media (unbuffered RPMI 1640, 10 mM glucose, 10% FCS) (Sigma-Aldrich) and allowed to adhere at 37 °C. Once adhered, 200 µl of additional media was added to each well, and the J774 cells were analyzed per the manufacturer's instructions to obtain real-time measurements of baseline OCRs and ECARs. Where indicated, ECARs and/or OCRs were analyzed in response to 1 μM oligomycin, 1.5 μM fluoro-carbonyl cyanide phenylhydrazone, and 100 nM rotenone plus 1 μM antimycin A (Sigma-Aldrich).
Hoyt L.R., Randall M.J., Ather J.L., DePuccio D.P., Landry C.C., Qian X., Janssen-Heininger Y.M., van der Vliet A., Dixon A.E., Amiel E, & Poynter M.E. (2017). Mitochondrial ROS induced by chronic ethanol exposure promote hyper-activation of the NLRP3 inflammasome. Redox Biology, 12, 883-896.
+ Open protocol
+ Expand
2

Mitochondrial Function Evaluation in C2C12 Myoblasts

Check if the same lab product or an alternative is used in the 5 most similar protocols
C2C12 myoblasts were seeded in XF 24-well microplates (Seahorse Bioscience, Billerica, USA) and differentiated. Mitochondrial inhibitors, including 1 μmol/L oligomycin, 1 μmol/L carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and 1 μmol/L rotenone/antimycin A, were incubated with these cells respectively. The cell OCR was measured by the extracellular flux analysis (Seahorse Biosciences) every 8 min.
Basal respiration represents the baseline oxygen consumption reading prior to the addition of inhibitors, antimycin A Rotenone, with the subtraction of non-mitochondrial respiration. Maximal respiration is shown as the maximum OCR measurement value following FCCP injection and with the subtraction of non-mitochondrial respiration. ATP production respiration represents the OCR reduction after the addition of an inhibitor of ATP synthase, oligomycin. Spare respiratory capacity is calculated by maximal respiration subtracting basal respiration. After detection, cell protein content was calculated, and OCR was adjusted accordingly.
Zhang W., You B., Qi D., Qiu L., Ripley-Gonzalez J.W., Zheng F., Fu S., Li C., Dun Y, & Liu S. (2021). Trimetazidine and exercise provide comparable improvements to high fat diet-induced muscle dysfunction through enhancement of mitochondrial quality control. Scientific Reports, 11, 19116.
+ Open protocol
+ Expand
3

Mitochondrial Metabolic Profiling by Seahorse

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Mitochondrial metabolic profiling was performed using extracellular flux analysis (Seahorse Bioscience/Agilent, Mulgrave, Australia) as described by us previously [105 (link)]. Basal, phosphorylating, maximal and non-mitochondrial respiration was measured initially through the sequential addition of 2 µg/mL oligomycin, 400 nm carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) with 10 mM sodium pyruvate, and 1 µM antimycin A, respectively. The extracellular acidification rate (ECAR) was concomitantly measured as an indicator of anaerobic glycolysis.
Campelj D.G., Timpani C.A., Petersen A.C., Hayes A., Goodman C.A, & Rybalka E. (2020). The Paradoxical Effect of PARP Inhibitor BGP-15 on Irinotecan-Induced Cachexia and Skeletal Muscle Dysfunction. Cancers, 12(12), 3810.
+ Open protocol
+ Expand
4

Mitochondrial Respiration in C2C12 Myoblasts

Check if the same lab product or an alternative is used in the 5 most similar protocols
C2C12 myoblasts were seeded in XF 24-well microplates (Seahorse Bioscience, Billerica, MA, USA). After differentiation and treatment, oxygen consumption was measured with extracellular flux analysis (Seahorse Biosciences). The final concentrations of mitochondrial inhibitors were at 10 μM antimycin A, 6 μM FCCP and 10 μM oligomycin. Basal respiration is the baseline oxygen consumption reading before compounds are injected. Maximal respiration represents the maximum OCR measurement value after FCCP injection. Spare respiratory capacity is calculated by noting the OCR response to FCCP, and dividing that number by the basal respiration to get a percentage. After detection, cells numbers were calculated and OCR was adjusted accordingly.
Wang X., Li H., Zheng A., Yang L., Liu J., Chen C., Tang Y., Zou X., Li Y., Long J., Liu J., Zhang Y, & Feng Z. (2014). Mitochondrial dysfunction-associated OPA1 cleavage contributes to muscle degeneration: preventative effect of hydroxytyrosol acetate. Cell Death & Disease, 5(11), e1521-.
+ Open protocol
+ Expand
5

Mitochondrial Function in C2C12 Myoblasts

Check if the same lab product or an alternative is used in the 5 most similar protocols
C2C12 myoblasts were seeded in XF 24-well microplates (Seahorse Bioscience, Billerica, MA, United States) and differentiated. Following treatments of mitochondrial inhibitors, including 1 μmol/L oligomycin, 1 μmol/L Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) and 1 μmol/L rotenone/antimycin A, OCRs were measured with extracellular flux analysis (Seahorse Biosciences) every 8 minutes.
You B., Dun Y., Fu S., Qi D., Zhang W., Liu Y., Qiu L., Xie M, & Liu S. (2021). The Treatment of Rhodiola Mimics Exercise to Resist High-Fat Diet-Induced Muscle Dysfunction via Sirtuin1-Dependent Mechanisms. Frontiers in Pharmacology, 12, 646489.
+ Open protocol
+ Expand
6

Extracellular Flux Analysis of IgA+ Plasma Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Extracellular flux analysis (Agilent, USA) was performed with replicates of 150,000 sort-purified IgA+ PCs isolated from small intestinal lamina propria or IgD+ B cells isolated from Peyer’s patches. Cells were adhered to each well of the Seahorse plate (Seahorse/Agilent, USA) using CellTak (Corning, USA). Cells were rested in Seahorse medium (Glucose-free DMEM) at 37°C without CO2 for at least 30 minutes prior to the run. For the test, Seahorse XF medium was supplemented with 2mM of L-glutamine (Sigma, UK) and pH was adjusted to 7.35±0.05 (at 37°C). Glucose (10mM final concentration) (Fischer Scientific, USA), oligomycin (1µM final concentration (Sigma, UK) and 2-DG (100mM final concentration; Sigma), were added to individual ports to complete this assay.
Penny H.A., Domingues R.G., Krauss M.Z., Melo-Gonzalez F., Lawson M.A., Dickson S., Parkinson J., Hurry M., Purse C., Jegham E., Godinho-Silva C., Rendas M., Veiga-Fernandes H., Bechtold D.A., Grencis R.K., Toellner K.M., Waisman A., Swann J.R., Gibbs J.E, & Hepworth M.R. (2022). Rhythmicity of Intestinal IgA Responses Confers Oscillatory Commensal Microbiota Mutualism. Science immunology, 7(75), eabk2541.
+ Open protocol
+ Expand
7

Extracellular Flux Analysis of IgA+ Plasma Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Extracellular flux analysis (Agilent, USA) was performed with replicates of 150,000 sort-purified IgA+ PCs isolated from small intestinal lamina propria or IgD+ B cells isolated from Peyer’s patches. Cells were adhered to each well of the Seahorse plate (Seahorse/Agilent, USA) using CellTak (Corning, USA). Cells were rested in Seahorse medium (Glucose-free DMEM) at 37°C without CO2 for at least 30 minutes prior to the run. For the test, Seahorse XF medium was supplemented with 2mM of L-glutamine (Sigma, UK) and pH was adjusted to 7.35±0.05 (at 37°C). Glucose (10mM final concentration) (Fischer Scientific, USA), oligomycin (1µM final concentration (Sigma, UK) and 2-DG (100mM final concentration; Sigma), were added to individual ports to complete this assay.
Penny H.A., Domingues R.G., Krauss M.Z., Melo-Gonzalez F., Lawson M.A., Dickson S., Parkinson J., Hurry M., Purse C., Jegham E., Godinho-Silva C., Rendas M., Veiga-Fernandes H., Bechtold D.A., Grencis R.K., Toellner K.M., Waisman A., Swann J.R., Gibbs J.E, & Hepworth M.R. (2022). Rhythmicity of Intestinal IgA Responses Confers Oscillatory Commensal Microbiota Mutualism. Science immunology, 7(75), eabk2541.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!