Complex I Enzyme Activity Dipstick Assay Kit is a dipstick assay designed to measure Complex I (mitochondria) activity in human, mouse, rat and bovine samples.
- 48 tests format
- Activity assay not sensitive to retonone for more specificity and simplicity
- Read on dipstick reader - use of standard imaging system also possible
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- Orphanet journal of rare diseases 18:802023Alpha-lipoic acid supplementation corrects pathological alterations in cellular models of pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMarta Talaverón-Rey,Mónica Álvarez-Córdoba,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,David Gómez-Fernández,Ana Romero-González,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Paula Cilleros-Holgado,Diana Reche-López,Rocío Piñero-Pérez,José A Sánchez-AlcázarPubMed 37046296
- Nature communications 14:11722023Transgenic NADH dehydrogenase restores oxygen regulation of breathing in mitochondrial complex I-deficient mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBlanca Jiménez-Gómez,Patricia Ortega-Sáenz,Lin Gao,Patricia González-Rodríguez,Paula García-Flores,Navdeep Chandel,José López-BarneoPubMed 36859533
- Redox biology 58:1025082022SARS-CoV-2 mitochondriopathy in COVID-19 pneumonia exacerbates hypoxemia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesStephen L Archer,Asish Dasgupta,Kuang-Hueih Chen,Danchen Wu,Kaushal Baid,John E Mamatis,Victoria Gonzalez,Austin Read,Rachel Et Bentley,Ashley Y Martin,Jeffrey D Mewburn,Kimberly J Dunham-Snary,Gerald A Evans,Gary Levy,Oliver Jones,Ruaa Al-Qazazi,Brooke Ring,Elahe Alizadeh,Charles Ct Hindmarch,Jenna Rossi,Patricia DA Lima,Darryl Falzarano,Arinjay Banerjee,Che C ColpittsPubMed 36334378
- Orphanet journal of rare diseases 17:3112022Therapeutic approach with commercial supplements for pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMónica Álvarez-Córdoba,Diana Reche-López,Paula Cilleros-Holgado,Marta Talaverón-Rey,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Rocío Piñero-Pérez,José A Sánchez-AlcázarPubMed 35945593
- Orphanet journal of rare diseases 17:2042022UPR activation improves pathological alterations in cellular models of mitochondrial diseases.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJuan M Suárez-Rivero,Carmen J Pastor-Maldonado,Suleva Povea-Cabello,Mónica Álvarez-Córdoba,Irene Villalón-García,Marta Talaverón-Rey,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Diana Reche-López,Paula Cilleros-Holgado,Rocío Piñero-Perez,José A Sánchez-AlcázarPubMed 35581596
- Frontiers in pharmacology 13:8620852022Pterostilbene in Combination With Mitochondrial Cofactors Improve Mitochondrial Function in Cellular Models of Mitochondrial Diseases.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJuan M Suárez-Rivero,Carmen J Pastor-Maldonado,Ana Romero-González,David Gómez-Fernandez,Suleva Povea-Cabello,Mónica Álvarez-Córdoba,Irene Villalón-García,Marta Talaverón-Rey,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,José A Sánchez-AlcázarPubMed 35370630
- Clinical genetics 102:56-602022A novel homozygous variant in COX5A causes an attenuated phenotype with failure to thrive, lactic acidosis, hypoglycemia, and short stature.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlessandra Torraco,Silvia Morlino,Teresa Rizza,Michela Di Nottia,Giorgia Bottaro,Luigi Bisceglia,Arianna Montanari,Marco Cappa,Marco Castori,Enrico Bertini,Rosalba CarrozzoPubMed 35246835
- The Prostate 81:1135-11482021SIRT3 affects mitochondrial metabolic reprogramming via the AMPK-PGC-1α axis in the development of benign prostatic hyperplasia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYongzhi Li,Qian Wang,Jingyu Li,Benkang Shi,Yili Liu,Ping WangPubMed 34411320
- Orphanet journal of rare diseases 16:2012021Down regulation of the expression of mitochondrial phosphopantetheinyl-proteins in pantothenate kinase-associated neurodegeneration: pathophysiological consequences and therapeutic perspectives.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMónica Álvarez-Córdoba,Marta Talaverón-Rey,Irene Villalón-García,Suleva Povea-Cabello,Juan M Suárez-Rivero,Alejandra Suárez-Carrillo,Manuel Munuera-Cabeza,Joaquín J Salas,José A Sánchez-AlcázarPubMed 33952316
- Scientific reports 10:170122020Simvastatin improves mitochondrial respiration in peripheral blood cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJon Ambæk Durhuus et. al.PubMed 33046789
Key facts
- Sample types
- Cell culture extracts, Tissue, Cell culture supernatant
- Assay type
- Enzyme activity
- Reactive species
- Mouse, Rat, Cow, Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application FuncS | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
As part of the MCIA complex, involved in the assembly of the mitochondrial complex I.
Additional Targets
NDUFAF2, AIFM1, NDUFS4, ECSIT, NDUFAF4, NDUFAF2
Alternative names
CIA30, CGI-65, NDUFAF1, NADH dehydrogenase [ubiquinone] 1 alpha subcomplex assembly factor 1
What's included?
Recommended products
Complex I Enzyme Activity Dipstick Assay Kit is a dipstick assay designed to measure Complex I (mitochondria) activity in human, mouse, rat and bovine samples.
- 48 tests format
- Activity assay not sensitive to retonone for more specificity and simplicity
- Read on dipstick reader - use of standard imaging system also possible
Key facts
- Sample types
- Cell culture extracts, Tissue, Cell culture supernatant
- Assay type
- Enzyme activity
- Reactive species
- Mouse, Rat, Cow, Human
- Assay Platform
- Reagents
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Contains 48 dipsticks and necessary components to quantify the activity of the Complex I enzyme complex in human, bovine, rat and mouse samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.
In this assay the specificity of anti-Complex I monoclonal antibodies (mAbs) is combined with the well-characterized Complex I in-gel activity assay that is not rotenone sensitive. First, Complex I is immunocaptured (i.e. immuno-precipitated in active form) on the dipstick. Second, the dipstick is immersed in Complex I activity buffer solution containing NADH as a substrate and nitrotetrazolium blue (NBT) as the electron acceptor. Immunocaptured Complex I oxidizes NADH and the resulting H+ reduces NBT to form a blue-purple precipitate at the Complex I antibody line on the dipstick. The signal intensity of this precipitate corresponds to the level of Complex I enzyme activity in the sample. Combined with dipstick assay kit for measuring Complex I quantity (Complex I Human Protein Quantity Dipstick Assay Kit ab109722/MS131 for human sample; ab109875/MS133 for rodent samples), it is possible to determine the relative specific activity of immunocaptured Complex I. The signal intensity is best measured by a dipstick reader or may be analyzed by a standard imaging system.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (ab109720)
An example using ab109720 to measure Complex I activity in human fibroblast samples. Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 30 μg of fibroblast protein extract.
Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (ab109720)
An example using ab109720 to measure Complex I activity in human fibroblast samples. Based on the standard curve, 15 μg of protein extract were loaded onto a dipstick for each sample. The figure shows four developed dipsticks, a control sample (1) and three unknowns (2-4). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 14-61% of normal Complex I activity levels.
Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (ab109720)
Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.
This image is courtesy of a customer review submitted by Christian MarxFunctional Studies - Complex I Enzyme Activity Dipstick Assay Kit (ab109720)
ab109720 was used to quantify complex I activity in HCT116 cells. The kit was used as described in the manual.
Shortly: cells were scraped and lysed in extraction buffer. Cell fragments were removed bey centrifugation and protein concentration was measured by Bradford assay. Increasing amounts of protein (as shown in image) were applied to each well as standard curve. A standard flatbed scanner was used instead of a Dipstick-reader. Due to the low contrast of the resulting bands, the brightness and contrast of the image was adjusted afterwards. Developed for 45 minutes as recommended by the manual.
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