Overview

  • Product nameACADM ELISA Kit
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 12 4.5%
    Overall 12 4.5%
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay typeSandwich (quantitative)
  • Sensitivity
    2 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 104 95% - 118%
    Cell culture media 105 97% - 114%

  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab129734 Medium-chain-acyl-CoA dehydrogenase (MCAD, gene ACADM) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the accurate quantitative measurement of medium-chain-acyl-CoA dehydrogenase in human, rat and mouse tissue and cell culture extracts. The assay employs an ACADM specific antibody coated onto well plate strips. Standards and samples are pipetted into the wells and ACADM present in the sample is bound to the wells by the immobilized antibody. The wells are washed and a primary anti-ACADM detector antibody is added. After washing away unbound detector antibody, an HRP-conjugated secondary detector antibody (HRP Label) specific for the primary detector antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ACADM bound. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.

    MCAD (medium-chain acyl-CoA dehydrogenase), encoded by the ACADM gene, is an oxidoreductase enzyme of the mitochondrial fatty acid beta-oxidation pathway that is specific for acyl chain lengths of 4 to 16. MCAD utilizes the electron transfer flavoprotein (ETF) as an electron acceptor that transfers the electrons to the main mitochondrial respiratory chain via ETF-ubiquinone oxidoreductase (ETF dehydrogenase).

  • Tested applicationsELISAmore details
  • PlatformMicroplate

Properties

  • RelevanceThe gene encodes the medium-chain specific (C4 to C12 straight chain) acyl-Coenzyme A dehydrogenase. The homotetramer enzyme catalyzes the initial step of the mitochondrial fatty acid beta-oxidation pathway. Defects in this gene cause medium-chain acyl-CoA dehydrogenase deficiency, a disease characterized by hepatic dysfunction, fasting hypoglycemia, and encephalopathy, which can result in infantile death. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.
  • Cellular localizationMitochondrion matrix.
  • Alternative names
    • ACAD1
    • Acyl-CoA dehydrogenase, C-4 to C-12 straight chain
    • Acyl-CoA dehydrogenase, medium chain
    • MCAD
    • MCADH
    • Medium-chain specific acyl-CoA dehydrogenase, mitochondrial
    • Medium-chain, Acyl-CoA dehydrogenase
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab129734 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
ELISA Use at an assay dependent concentration.

ACADM ELISA Kit images

  • Example of serially titrated ACADM protein in the assay working range.
  • The graph above shows an experiment to determine the levels of ACADM encoded protein in various fibroblast cell lines. Three Medium Chain Acyl-CoA Dehydrogenase Deficient (MCADD) patient fibroblast cell lines (P1-3) each harboring the common MCAD mutation K304E (984A>G in ACADM gene) were analyzed. Each patient cell line shows significant reduction in ACADM protein expression when compared to control adult fibroblast cells from three unaffected individuals (CON1-3).

Protocols

References for ACADM ELISA Kit (ab129734)

This product has been referenced in:
  • Koster KL  et al. Functional studies of 18 heterologously expressed medium-chain acyl-CoA dehydrogenase (MCAD) variants. J Inherit Metab Dis N/A:N/A (2014). ELISA ; Human . Read more (PubMed: 24966162) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"