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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.

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Functional Studies - MitoTox™ Complex II OXPHOS Activity Assay Kit (AB109904), expandable thumbnail

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Key facts

Detection method

Colorimetric

Assay type

Direct

Reactive species

Mouse, Cow, Human

Assay time

4h

Associated Products

Select an associated product type

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Target data

Function

Polynucleotide kinase that can phosphorylate the 5'-hydroxyl groups of double-stranded RNA (dsRNA), single-stranded RNA (ssRNA), double-stranded DNA (dsDNA) and double-stranded DNA:RNA hybrids. dsRNA is phosphorylated more efficiently than dsDNA, and the RNA component of a DNA:RNA hybrid is phosphorylated more efficiently than the DNA component. Plays a key role in both tRNA splicing and mRNA 3'-end formation. Component of the tRNA splicing endonuclease complex: phosphorylates the 5'-terminus of the tRNA 3'-exon during tRNA splicing; this phosphorylation event is a prerequisite for the subsequent ligation of the two exon halves and the production of a mature tRNA (PubMed:24766809, PubMed:24766810). Its role in tRNA splicing and maturation is required for cerebellar development (PubMed:24766809, PubMed:24766810). Component of the pre-mRNA cleavage complex II (CF-II), which seems to be required for mRNA 3'-end formation. Also phosphorylates the 5'-terminus of exogenously introduced short interfering RNAs (siRNAs), which is a necessary prerequisite for their incorporation into the RNA-induced silencing complex (RISC). However, endogenous siRNAs and microRNAs (miRNAs) that are produced by the cleavage of dsRNA precursors by DICER1 already contain a 5'-phosphate group, so this protein may be dispensible for normal RNA-mediated gene silencing.

Targets

SDHA

Alternative names

Polyribonucleotide 5'-hydroxyl-kinase Clp1, Polyadenylation factor Clp1, Polynucleotide kinase Clp1, Pre-mRNA cleavage complex II protein Clp1, HEAB, CLP1

What's included?

96 Test
Components
12-channel reagent reservoirs
2 x 1 Unit
20X Buffer
1 x 5 mL
Bovine heart mitochondria
2 x 360 µL
Complex II Activity Buffer
1 x 25 mL
DCPIP/DCIP
1 x 250 µL
Detergent
1 x 1 mL
Pre-coated 96-well microplate
1 x 96 Test
Succinate
1 x 500 µL
Ubiquinone 2
1 x 60 µL

Recommended products

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.

Key facts

Detection method

Colorimetric

Assay type

Direct

Reactive species

Mouse, Cow, Human

Assay time

4h

Assay Platform

Microplate reader

Storage

Shipped at conditions

Dry Ice

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

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. **Related products** Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Complex II also known as succinate dehydrogenase (SDH) or succinate-ubiquinone oxidoreductase plays an important mechanical role in the mitochondrial electron transport chain. It catalyzes the oxidation of succinate to fumarate while reducing ubiquinone to ubiquinol. The enzyme complex is approximately 140 kDa in mass and resides in the inner mitochondrial membrane. Complex II is expressed in most tissues particularly in high-energy demand tissues such as the heart and skeletal muscles.

Biological function summary

Complex II functions as part of the larger electron transport chain complex and plays a role in the Krebs cycle. It links two critical metabolic pathways converting succinate to fumarate while transferring electrons to the electron transport chain. This makes it integral for proper cellular respiration and energy production. The complex consists of multiple subunits and utilizes co-factors like FAD and iron-sulfur clusters for enzymatic activity. It is also a part of the supercomplexes that optimize the efficiency of oxidative phosphorylation.

Pathways

Complex II plays a significant role in both the citric acid cycle and oxidative phosphorylation. It acts as a connecting bridge between these two pathways facilitating the flow of electrons. Complex II works alongside other proteins such as complex I and complex III to maintain the electron transport chain's function and energy production. Succinate dehydrogenase transfers electrons within the chain directly affecting the generation of ATP by complex V (ATP synthase).

Associated diseases and disorders

Dysfunction of complex II is associated with mitochondrial diseases and cancers. Mutations or deficiencies in its subunits can lead to conditions like Leigh syndrome and hereditary paraganglioma. These conditions frequently involve other mitochondrial proteins and complexes such as complex I which can exacerbate the electron transport chain dysfunction. In cancers alterations in succinate dehydrogenase activity can result in oncogenic metabolisms by falsely stabilizing hypoxia-inducible factors linking it further with the genetic and metabolic regulation.

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  • Functional Studies - MitoTox™ Complex II OXPHOS Activity Assay Kit (ab109904), expandable thumbnail

    Functional Studies - MitoTox™ Complex II OXPHOS Activity Assay Kit (ab109904)

    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.

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Product protocols

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