Key features and details
- One-wash 90 minute protocol
- Sensitivity: 430 ng/ml
- Range: 3.13 µg/ml - 200 µg/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman NADH dehydrogenase ELISA Kit (Complex I)
See all Complex I kits
Intra-assay Sample n Mean SD CV% HepG2 lysate 5 2.7% Inter-assay Sample n Mean SD CV% HepG2 lysate 3 5.9%
Sample typeCell culture extracts, Tissue Extracts
Assay typeSandwich (quantitative)
Range3.13 µg/ml - 200 µg/ml
Sample specific recovery Sample type Average % Range Serum 110 108% - 111% Cell culture media 66 64% - 67% Fetal Bovine Serum 74 70% - 77%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Human NADH Dehydrogenase (Complex I) ELISA kit (ab178011) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of NADH Dehydrogenase protein in human cell and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human NADH Dehydrogenase with 430 ng/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.
NADH dehydrogenase (NADH: ubiquinone reductase (H+-translocating), Complex I) is the first enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. NADH dehydrogenase is a large protein complex of 950,000 MW made up of 45-46 different subunits. Seven of the subunits of the complex are encoded on mitochondrial DNA (mtDNA), the remaining subunits are nuclear encoded, made in the cytosol and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from NADH via a flavin (FMN) and several non-heme iron centers. Mutations in mtDNA, or nuclear DNA genes encoding NADH dehydrogenase subunits or assembly factors are a common cause of genetic OXPHOS defects. Mutations or loss of mtDNA may cause enzymatic dysfunction by disrupting enzyme assembly or alternatively by specifically affecting enzymatic activity with no effect on enzyme assembly.
NADH dehydrogenase (like Complex III) has been proposed as a site of superoxide ‘leak’ from the mitochondrial OXPHOS system. Altered functioning and increased superoxide production by this complex has been proposed to contribute to several neurological disorders including Parkinson's disease. Also there is evidence of NADH Dehydrogenase involvement in diabetes.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Human NADH Dehydrogenase Capture Antibody 1 x 600µl 10X Human NADH Dehydrogenase Detector Antibody 1 x 600µl 10X Wash Buffer LM 1 x 20ml 2X Cell Extraction Buffer LM 1 x 10ml Antibody Diluent 5B 1 x 6ml HeLa Human Lyophilized Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
- NADH dehydrogenase
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Example of human NADH Dehydrogenase control curve in 1X Cell Extraction Buffer LM. The NADH Dehydrogenase control curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Background-subtracted data values from triplicate measurements (mean +/- SD) are graphed.
Background-subtracted data values from triplicate measurements of three lysate concentrations (200, 100 and 50 µg/mL) are graphed as mean +/- SD.
Quantification of NADH Dehydrogenase expression in 143B wildtype (WT) and two clones (3B10 and 3F5) of 143B-derived Rho0 (mitochondrial DNA-depleted) cells. The concentrations of NADH Dehydrogenase were interpolated from data values shown in Figure 3 using NADH Dehydrogenase control curve of the HeLa Lyophilized Lysate Control, corrected for sample dilution, and graphed in percent relative to NADH Dehydrogenase expression in HeLa cell extract. The concentration of NADH Dehydrogenase in both Rho0 cell lines was less than 1% of the concentration in the WT 143B cells.
ab178011 has been referenced in 7 publications.
- Kuang Y et al. NADH dehydrogenase subunit 1/4/5 promotes survival of acute myeloid leukemia by mediating specific oxidative phosphorylation. Mol Med Rep 25:N/A (2022). PubMed: 35425997
- Tiwari R et al. Chemical inhibition of oxygen-sensing prolyl hydroxylases impairs angiogenic competence of human vascular endothelium through metabolic reprogramming. iScience 25:105086 (2022). PubMed: 36157579
- Lombardi M et al. Mitochondrial Energetics and Ca2+-Activated ATPase in Obstructive Hypertrophic Cardiomyopathy. J Clin Med 9:N/A (2020). PubMed: 32527005
- Jaworski S et al. Degradation of Mitochondria and Oxidative Stress as the Main Mechanism of Toxicity of Pristine Graphene on U87 Glioblastoma Cells and Tumors and HS-5 Cells. Int J Mol Sci 20:N/A (2019). PubMed: 30717385
- Khasawneh RR et al. Cross talk between 26S proteasome and mitochondria in human mesenchymal stem cells' ability to survive under hypoxia stress. J Physiol Sci 69:1005-1017 (2019). PubMed: 31679117
- Andreux PA et al. Mitochondrial function is impaired in the skeletal muscle of pre-frail elderly. Sci Rep 8:8548 (2018). PubMed: 29867098
- Rollins BL et al. Mitochondrial Complex I Deficiency in Schizophrenia and Bipolar Disorder and Medication Influence. Mol Neuropsychiatry 3:157-169 (2018). PubMed: 29594135