MitoTox™ Complex I OXPHOS Activity Assay Kit (ab109903)
- Datasheet
- Specific References (5)
- Protocols
Overview
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Product nameMitoTox™ Complex I OXPHOS Activity Assay Kit
See all Complex I kits -
Detection methodColorimetric
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Sample typeInhibitor compounds
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Assay typeQuantitative
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Assay time5h 00m
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Product overview
MitoTox™ Complex I OXPHOS Activity Assay (ab109903) is designed for testing the direct inhibitory effect of compounds on Complex I activity in only 5 hours. Complex I extracted from the provided bovine heart mitochondria (a rich source of Complex I) is immunocaptured by specific antibodies on the plate. Complex I activity can be observed as decrease in absorbance at OD 340 nm. The intra-assay and inter-assay variation of this assay are both < 10%.
Inhibitory effects of compounds on Complex I activity can be tested in two different ways: 1. Screening format, where up to 23 compounds can be tested at a single concentration in triplicate; 2. Dose response (IC50) format, where two compounds known to affect Complex I activity can be tested at 11 different data points in triplicate.
Testing for mitochondrial function has become a key aspect of drug discovery. Mitochondria can be affected by drug treatment, resulting into cardio- and hepatotoxic side effects that can lead to drug withdrawal from the market. Therefore, there is increasing emphasis on testing the impact on mitochondria early on in the drug development process to reduce failure rates during preclinical and clinical phases.
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Notes
Store Phospholipids, Bovine heart mitochondria, Ubiquinone and Activity Buffer at -80°C. Store all other components at 4°C.
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PlatformMicroplate
Properties
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Storage instructionsPlease refer to protocols.
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Components 96 tests 12-channel reagent reservoirs 2 units 1X Mito Buffer 1 x 5ml 20X Wash Buffer 1 x 5ml Bovine heart mitochondria 1 x 360µl Complex I Activity Buffer 1 x 24ml Detergent 1 x 100µl Phospholipids 1 x 6ml Pre-coated 96-well microplate 1 unit Ubiquinone 1 1 x 60µl -
Research areas
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Alternative names
- NADH dehydrogenase
Images
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MitoTox™ Complex I OXPHOS Activity Assay (ab109903)Typical dose response curve for rotenone. Assay was performed following the Dose Response Assay Procedure using rotenone, a well known Complex I inhibitor. Rotenone was prepared in DMSO to generate a 10 mM stock. Starting with a 50 µM final concentration in well, 1:10 serial dilutions of rotenone were generated.
Protocols
References
This product has been referenced in:
- Cameron AR et al. Metformin selectively targets redox control of complex I energy transduction. Redox Biol 14:187-197 (2018). Read more (PubMed: 28942196) »
- Choi SE et al. NF-?B/p53-activated inflammatory response involves in diquat-induced mitochondrial dysfunction and apoptosis. Environ Toxicol N/A:N/A (2018). Read more (PubMed: 29484840) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"