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Product name
MitoTox™ Complex III OXPHOS Activity Microplate Assay
See all Complex III products (3) ...
Tests
1 x 96 well plate
Assay type
Direct
Assay time
0h 30m
Cross reactivity
Reacts with
Cow
Product overview
The MitoTox™ OXPHOS Complex III Activity Kit is designed for screening the direct inhibitory effects of compounds on Complex II and III activity. Bovine heart mitochondria are provided in this kit as they are a rich source of these two enzymes. The kit contains succinate (Solution 1) which is the electron donor of Complex II, as well as cytochrome c (oxidized), the electron acceptor of Complex III. Ubiquinone, the electron acceptor of Complex II, is present in the mitochondria. The rate of the coupled Complex II+III reaction is measured by monitoring the conversion of cytochrome c in its oxidized form, cytochrome c (oxidized), to its reduced form, cytochrome c (reduced), as a linear increase in absorbance at 550 nm. The kit requires the use of rotenone and KCN (not supplied with the kit). Addition of rotenone, an inhibitor of Complex I, ensures that all reduction of Cytochrome c is via Complex II. Addition of KCN ensures that there is no re-oxidation of Cytochrome c by Complex IV. The user may add antimycin, an inhibitor of Complex III activity, in the screening procedure as a positive control. 50% inhibition of Complex III activity is obtained with 22 ± 4 nM antimycin at assay conditions described in the protocol.
Notes
Store at -80°C: Bovine Heart Mitochondria, Cytochrome c (oxidized).
Store at 4°C: Succinate Solution, Microplate, Mito Buffer and Reagent Reservoirs: Reagent Reservoirs may be stored at room temperature.
Store at 4°C: Succinate Solution, Microplate, Mito Buffer and Reagent Reservoirs: Reagent Reservoirs may be stored at room temperature.
Tested applications
Functional Studiesmore details
Properties
Storage instructions
Please see Notes section
| Components | 96 tests |
|---|---|
| 12-channel reagent reservoirs | 2 units |
| 1X Complex III Mito Buffer | 1 x 1ml |
| 1X Succinate Solution | 1 x 12ml |
| 96-well microplate | 1 unit |
| Bovine heart mitochondria (5 mg/mL) | 1 x 300µl |
| Cytochrome c (oxidized) | 1 x 550µl |
| Single-channel reagent reservoirs | 2 units |
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Functional Studies - MitoTox™ Complex III OXPHOS Activity Microplate Assay (ab109905)
(enlarge)
Relevance
Complex II (succinate-ubiquinone oxidoreductase), one of the mitochondrial respiratory chain complexes, transfers electrons from succinate generated during the citric acid cycle to Complex III (ubiquinolcytochrome c oxidoreductase), via a mobile electron shuttle, ubiquinone. Complex III transfers electrons to Complex IV (cytochrome c oxidase) via another mobile electron shuttle, cytochrome c.
Alternative names
- MS1037
- MTOX3
Applications
Show applications key
Our Abpromise guarantee covers the use of ab109905 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
FuncS
MitoTox™ Complex III OXPHOS Activity Microplate Assay images:
Protocols
References for MitoTox™ Complex III OXPHOS Activity Microplate Assay (ab109905)
ab109905 has not yet been referenced specifically in any publications.
Publishing research using ab109905? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"