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A simple, plate-based colorimetric assay for measuring ATP synthase in isolated mitochondria
MILPITAS, Calif. - Californer -- Mitochondrial ATP synthase (EC 3.6.3.14), also known as mitochondrial complex V or F1F0-ATP synthase is a multi-subunit membrane-bound enzyme of mitochondria that utilizes the proton electrochemical gradient generated by the respiratory chain for ATP synthesis. It consists of two domains, an inner membrane embedded domain (F0) and a membrane extrinsic domain (F1), which is also the catalytic core of the enzyme.
In addition to its critical role in ATP synthesis, ATP synthase regulates the flux of oxidative phosphorylation, mitochondrial signaling by reactive oxygen species, cell death etc. Defects in ATP synthase have been associated with human diseases including neuromuscular disorders, heart failure etc. Thus, ATP synthase may be used as a therapeutic drug target in the treatment of many diseases.
BioVision's Mitochondrial ATP Synthase Activity Assay Kit is a simple, plate-based colorimetric assay for measuring ATP synthase in isolated mitochondrial samples. In this assay, ATP Synthase hydrolyzes ATP to ADP. In the subsequent steps, ADP together with ATP synthase converter and enzyme mix oxidizes NADH to NAD+, which is monitored via a decrease in absorbance at 340 nm. A specific ATP synthase inhibitor, oligomycin is also included in the kit.
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Sample specific ATP synthase activity is obtained by measuring the sample ATP synthase activity in the presence and absence of oligomycin. The assay provides a simple, rapid, and sensitive method to measure ATP synthase activity in isolated mitochondria.
Figures. (A) ATP Synthase activity in purified mitochondria with and without Oligomycin.
(B) ATP Synthase activity using varying concentrations of Jurkat cell mitochondria (isolated using BioVision Cat# K288-50). (C) ATP Synthase activity in mitochondria isolated from Jurkat cells, HepG2 cells, rat heart (isolated using BioVision Cat# (K288-50) and bovine heart mitochondria (obtained commercially) with and without Oligomycin. Note: Net ATP Synthase activity in sample = Activity in reaction without Oligomycin – Activity in reaction with Oligomycin. All assays were performed following kit protocol.
Storage Conditions Multiple Temperatures
USAGE For Research Use Only! Not For Use in Humans.
For more information on this Assay Kit Cat# K2112 https://bit.ly/30Lsx5H
In addition to its critical role in ATP synthesis, ATP synthase regulates the flux of oxidative phosphorylation, mitochondrial signaling by reactive oxygen species, cell death etc. Defects in ATP synthase have been associated with human diseases including neuromuscular disorders, heart failure etc. Thus, ATP synthase may be used as a therapeutic drug target in the treatment of many diseases.
BioVision's Mitochondrial ATP Synthase Activity Assay Kit is a simple, plate-based colorimetric assay for measuring ATP synthase in isolated mitochondrial samples. In this assay, ATP Synthase hydrolyzes ATP to ADP. In the subsequent steps, ADP together with ATP synthase converter and enzyme mix oxidizes NADH to NAD+, which is monitored via a decrease in absorbance at 340 nm. A specific ATP synthase inhibitor, oligomycin is also included in the kit.
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Sample specific ATP synthase activity is obtained by measuring the sample ATP synthase activity in the presence and absence of oligomycin. The assay provides a simple, rapid, and sensitive method to measure ATP synthase activity in isolated mitochondria.
Figures. (A) ATP Synthase activity in purified mitochondria with and without Oligomycin.
(B) ATP Synthase activity using varying concentrations of Jurkat cell mitochondria (isolated using BioVision Cat# K288-50). (C) ATP Synthase activity in mitochondria isolated from Jurkat cells, HepG2 cells, rat heart (isolated using BioVision Cat# (K288-50) and bovine heart mitochondria (obtained commercially) with and without Oligomycin. Note: Net ATP Synthase activity in sample = Activity in reaction without Oligomycin – Activity in reaction with Oligomycin. All assays were performed following kit protocol.
Storage Conditions Multiple Temperatures
USAGE For Research Use Only! Not For Use in Humans.
For more information on this Assay Kit Cat# K2112 https://bit.ly/30Lsx5H
Source: BioVision Inc
Filed Under: Biotech
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