Overview

  • Product name

    Human NADH dehydrogenase ELISA Kit (Complex I)
    See all Complex I kits
  • Detection method

    Colorimetric
  • Precision

    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 type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.43 µg/ml
  • Range

    3.13 µg/ml - 200 µg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 110 108% - 111%
    Cell culture media 66 64% - 67%
    Fetal Bovine Serum 74 70% - 77%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s NADH Dehydrogenase (Complex I) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of NADH dehydrogenase protein in human cell and tissue extracts.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm.Optionally,instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab178011 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • The NADH Dehydrogenase standard curve was prepared using HeLa cell extract. 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.

  • The concentrations of NADH Dehydrogenase were interpolated from data values shown in the figure above using NADH Dehydrogenase standard curve of HeLa cell extract, 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.

Protocols

References

This product has been referenced in:

  • 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). Read more (PubMed: 30717385) »
  • Andreux PA  et al. Mitochondrial function is impaired in the skeletal muscle of pre-frail elderly. Sci Rep 8:8548 (2018). Read more (PubMed: 29867098) »
See all 3 Publications for this product

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