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    products/assay-kits/malate-dehydrogenase-1-mdh1-activity-assay-ab200009.pdf

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Signal Transduction Metabolism Energy Metabolism
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Malate Dehydrogenase 1 (MDH1) Activity Assay (ab200009)

  • Datasheet
  • SDS
  • Protocol Booklet
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Raw data from HeLa cell lysate.
  • Representative background-subtracted kinetic measurements
  • MDH1 activity in cell lysates and tissue homogenates
  • MDH1 activity in serially titrated HeLa cell fractions

Key features and details

  • Assay type: Enzyme activity (quantitative)
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: Cell culture extracts, Tissue Extracts

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Overview

  • Product name

    Malate Dehydrogenase 1 (MDH1) Activity Assay
    See all MDH1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    overall 4 3.4%
    Inter-assay
    Sample n Mean SD CV%
    overall 4 8.1%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Enzyme activity (quantitative)
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    Abcam’s Malate Dehydrogenase 1 (MDH1) Activity Assay kit is designed for the sensitive and accurate measurement of MDH1 activity in Human, mouse, and rat samples.

  • Notes

    Malate Dehydrogenase 1 is the cytoplasmic isoform of the enzyme responsible for catalyzing the reversible oxidation of malate to oxaloacetate. The enzyme’s activity requires the NAD+/NADH cofactor and participates primarily in the malate-aspartate shuttle.

    The enzyme activity is determined by following the production of NADH in the following MDH1 catalyzed reaction:

    Malate + NAD+ ↔ oxaloacetic acid + NADH

    The generation of NADH is coupled to the 1:1 reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450nm (Dye molar extinction coefficient: 37000M-1 cm-1). In each well, only native MDH1 is immunocaptured from the sample; this removes all other enzymes, including MDH2 from the activity measurement.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Coupler 1 vial
    100X NAD+ 1 vial
    100X Reagent Dye 1 vial
    100X Sodium Malate 1 vial
    10X Blocking Buffer 1 x 8ml
    20X Buffer 1 x 20ml
    Base Buffer 1 x 24ml
    Extraction Buffer (ab260490) 1 x 15ml
    Malate Dehydrogenase 1 Activity Microplate 1 unit
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
  • Sequence similarities

    Belongs to the LDH/MDH superfamily. MDH type 2 family.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P40925 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • cytoplasmic
    • Cytosolic malate dehydrogenase
    • Diiodophenylpyruvate reductase
    • Malate dehydrogenase
    • Malate dehydrogenase 1, NAD (soluble)
    • Malate dehydrogenase cytoplasmic
    • MDH s
    • mdh1
    • MDHA
    • MDHC_HUMAN
    • MDHs
    • MGC:1375
    • MOR2
    • Soluble malate dehydrogenase
    see all
  • Database links

    • Entrez Gene: 4190 Human
    • Entrez Gene: 17449 Mouse
    • Entrez Gene: 24551 Rat
    • Omim: 154200 Human
    • SwissProt: P40925 Human
    • SwissProt: P14152 Mouse
    • SwissProt: O88989 Rat
    • Unigene: 526521 Human
    • Unigene: 212703 Mouse
    • Unigene: 13492 Rat
    see all

Images

  • Raw data from HeLa cell lysate.
    Raw data from HeLa cell lysate.

    Once the rate/slope of each lane is extracted from the linear range of the time point data, it is expressed as rate (mOD/min) per microgram of cell lysate added per well.

  • Representative background-subtracted kinetic measurements
    Representative background-subtracted kinetic measurements

    Representative background-subtracted kinetic measurements from serially diluted HeLa cell lysates and tissue homogenates HLH, MLH, RLH.

  • MDH1 activity in cell lysates and tissue homogenates
    MDH1 activity in cell lysates and tissue homogenates

    The assay was used to determine the MDH1 activity in a series of normal cell lysates and tissue homogenates loaded at 250 µg/mL.

  • MDH1 activity in serially titrated HeLa cell fractions
    MDH1 activity in serially titrated HeLa cell fractions

    MDH1 activity in serially titrated HeLa cell fractions (Cyto = Cytoplasmic, Mito = Mitochondrial and Nuc = Nuclear) demonstrates the assay’s specificity to MDH1.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1)

Publishing research using ab200009? Please let us know so that we can cite the reference in this datasheet.

ab200009 has been referenced in 1 publication.

  • Wild J  et al. Aestivation motifs explain hypertension and muscle mass loss in mice with psoriatic skin barrier defect. Acta Physiol (Oxf) 232:e13628 (2021). PubMed: 33590724

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