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MitoTox™ Complex II OXPHOS Activity Assay Kit (ab109904) is designed for testing the direct inhibitory effect of compounds on Complex II activity in only 4 hours. Complex II extracted from the provided bovine heart mitochondria (a rich source of Complex II) is immunocaptured by specific antibodies on the plate. Complex II activity can be observed as decrease in absorbance at OD 600 nm. The intra-assay and inter-assay variation of this assay are both <15%.
Inhibitory effects of compounds on Complex II 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 II 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.
Please store Succinate, Ubiquinone 2, Bovine Heart Mitochondria, DCPIP at -80°C and all other components at 4°C.
|10X Detergent||1 x 1ml|
|12-channel reagent reservoirs||2 units|
|20 x Buffer||1 x 5ml|
|Bovine heart mitochondria||2 x 360µl|
|Complex II Activity Buffer||1 x 25ml|
|DCIP||1 x 250µl|
|Pre-coated 96-well microplate||1 x 96 tests|
|Succinate||1 x 500µl|
|Ubiquinone 2||1 x 60µl|
Typical dose response curve for TTFA (2-thenoyltrifluoracetone). Assay was performed following the Dose Response Assay Procedure using TTFA, a well known Complex II inhibitor. TTFA was prepared in DMSO to generate a 100 mM stock. Starting with a 500 µM final concentration in well, 1:2 serial dilutions of TTFA were generated.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"