Description
Key facts
Sample types:Cell culture extracts, Tissue,
Reacts with:Cow, Human,
Assay type:Sandwich (quantitative),
Assay Platform:Reagents
Product details:
ab109722 contains 48 dipsticks and necessary components to quantify the levels of the fully assembled Complex I enzyme complex in human and bovine samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.
Based on the immunologic sandwich assay, the kit utilizes two monoclonal antibodies (mAbs) specific to different antigens present on the Complex I enzyme complex. One antibody is immobilized on the nitrocellulose membrane in a thin line perpendicular to the length of the dipstick, while the other is gold-conjugated and combined with the sample mix. The sample contents containing the gold-conjugated mAb wick past the mAb immobilized on the dipstick. When assembled Complex I is present in the sample, a red line appears at the site of the anti-Complex I antibody line. The signal intensity is directly related to the amount of Complex I in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system. To identify defects in Complex I that do not affect enzyme assembly combine this assay with
ab109720
to determine the relative specific activity of Complex I.
All components are stable in their provided containers at room temperature out of direct sunlight.
After diluting the 10X Blocking Buffer to 2X, store at 4°C. For long-term storage, all buffers can be stored at 4°C.
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.
