Malate Dehydrogenase 1 (MDH1) Activity Assay (ab200009)

Overview

  • Product name

    Malate Dehydrogenase 1 (MDH1) Activity Assay
  • 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 1 x 15ml
    Malate Dehydrogenase 1 Activity Microplate 1 unit
  • Sequence similarities

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

    ISGylated.
    Acetylation at Lys-118 dramatically enhances enzymatic activity and promotes adipogenic differentiation.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Alternative names

    • cytoplasmic
    • Cytosolic malate dehydrogenase
    • Diiodophenylpyruvate reductase
    • Malate dehydrogenase
    • mdh1
    • MDHC_HUMAN
    see all
  • Database links

Images

  • 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 from serially diluted HeLa cell lysates and tissue homogenates HLH, MLH, RLH.

  • 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 (Cyto = Cytoplasmic, Mito = Mitochondrial and Nuc = Nuclear) demonstrates the assay’s specificity to MDH1.

Protocols

References

ab200009 has not yet been referenced specifically in any publications.

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