Description
Mitochondrial Complex I Activity Colorimetric Assay Kit
Mitochondrial Complex I or NADH:ubiquinone oxidoreductase (EC 1.6.5.3) is the first and the largest complex of the Electron Transport Chain located in the mitochondrial membrane. It oxidizes NADH to NAD+ and transfers an electron to ubiquinone (also present in the inner mitochondrial membrane) converting it to ubiquinol. During this process, it transports protons across the inner mitochondrial membrane, helping to develop an electrochemical gradient. This process is very important for cellular respiration and adverse effects on Complex I activity can compromise mitochondrial respiration, which further leads to cellular stress.
Assay Genie's Mitochondrial Complex I assay kit is a fast and reliable method to determine the activity of complex I in isolated mitochondria. It is useful for respiration studies in isolated mitochondria and may be used to study effects of toxicants, drugs and other environmental conditions on mitochondrial complex I activity. This kit uses decylubiquinone, an analog of ubiquinone, as an electron acceptor that gets converted to decylubiquinol through the catalytic activity of Complex I. The Complex I dye that absorbs light at 600 nm in its oxidized form is used as a terminal electron acceptor that accepts electrons from decylubiquinol. Complex I activity is determined colorimetrically by recording the change in absorbance of reduced Complex I dye at 600 nm. Specific Complex I activity is obtained by subtracting the activity in presence of Complex I inhibitor rotenone from total activity. This kit can detect as low as 0.1 mU / well and is linear up to 7 mU / well.
Figure: (a) Standard curve for oxidized Complex I dye. (b) OD600 with varying concentrations of Bovine heart mitochondria obtained commercially. (c) Complex I Activity in isolated Bovine heart mitochondria (obtained commercially) and jurkat cell mitochondria (isolated using cat # BN00546) with and without Complex I inhibitor rotenone.
New Column | New Column |
SKU | BN01128 |
Number of Assays | 100 assays |
Measurement | Colorimetric (Absorbance 600 nm) |
Reactivity | Multiple species |
Kit Overview | Measurement of Complex I activity in isolated mitochondria |
Features & Benefits | Simple, rapid & convenient assay to measure Mitochondrial Complex I Includes Inhibitor (Rotenone) |
Kit Contents |
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Storage Storage | -20°C |
Shipping Conditions | Gel Pack |
Use | For Research Use Only! Not For Use in Humans |