Description
Key facts
Reacts with:Mouse, Rat, Cow, Human,
Assay type:Quantitative
Product details:
Optimized for isolation of Complex I from small amounts of tissue for activity assays and other analyses. Based on a highly specific antibody.
250 μg of monoclonal antibody irreversibly crosslinked to protein G-agarose beads which can immunocapture ~60 μg of Complex I from heart mitochondria.
ab109711 allows for isolation of Complex I from small amounts of tissue. This facilitates subsequent analysis of assembly state and activity. Thus the enzyme retains NADH-ferricyanide, NADH-CoQ1 and NADH hexaaminoruthenium reductase activities after isolation, and with added lipids it also shows significant rotenone sensitivity. Finally, the extent of post translational modifications including oxidative damage can be readily analyzed by proteomic approaches or antibody detection of these modifications. Uses for ab109711 include but are not limited to examining alterations of Complex I subunits in inherited mitochondrial diseases, Parkinson's disease, Alzheimer's disease, ALS, schizophrenia, and aging.
Note: The immunocapture protocol for this kit requires Abcam detergent (lauryl maltoside,
ab109857
Product supplied as a 6.25% slurry. This consists of 250 μg antibody conjugated to 25 μL beads in a volume of 250 μL.
Related products
Review the
mitochondrial assay guide
, or the full
metabolism assay guide
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Complex I also known as NADH dehydrogenase or NADH:ubiquinone oxidoreductase is a large enzyme complex with a molecular mass of approximately 1000 kDa. It is expressed in the inner mitochondrial membrane of eukaryotic cells. As the first enzyme in the mitochondrial respiratory chain Complex I plays a critical mechanical role in cellular respiration. It transfers electrons from NADH to ubiquinone coupled with the translocation of protons across the inner mitochondrial membrane contributing to the generation of a proton gradient used to produce ATP.
Biological function summary
Complex I acts as an integral component of the mitochondrial respiratory chain which is a series of protein complexes involved in cellular energy production. As part of this complex system Complex I is essential for effective oxidative phosphorylation. Its activity is assessed using protein activity assays including immunocapture or complex activity assays and microplate assays. Complex I activity influences the overall efficiency of ATP production affecting energy-dependent cellular processes.
Pathways
Complex I functions within the electron transport chain one of the major pathways in cellular respiration. This pathway is vital for ATP synthesis providing the energy currency required by cells. Complex I works closely with other electron transport chain complexes such as Complex II (succinate dehydrogenase complex) and Complex III (cytochrome c reductase) to drive oxidation-reduction reactions and maintain cellular metabolism.
Defects in Complex I are linked with mitochondrial diseases and neurodegenerative disorders such as Leigh syndrome and Parkinson's disease. Mutations in Complex I subunits disrupt normal electron transport and ATP production leading to increased oxidative stress and neuronal cell damage. Other mitochondrial proteins such as cytochrome c oxidase are also implicated in these conditions emphasizing the interconnected nature of mitochondrial dysfunction in disease progression.
