Malate Dehydrogenase 1 (MDH1) Activity Assay (ab200009)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell culture extracts, Tissue Extracts
Overview
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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.
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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.
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Platform
Microplate reader
Properties
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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
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Sequence similarities
Belongs to the LDH/MDH superfamily. MDH type 2 family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- cytoplasmic
- Cytosolic malate dehydrogenase
- Diiodophenylpyruvate reductase
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
see all
Images
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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.
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Representative background-subtracted kinetic measurementsRepresentative background-subtracted kinetic measurements from serially diluted HeLa cell lysates and tissue homogenates HLH, MLH, RLH.
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MDH1 activity in cell lysates and tissue homogenatesThe assay was used to determine the MDH1 activity in a series of normal cell lysates and tissue homogenates loaded at 250 µg/mL.
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MDH1 activity in serially titrated HeLa cell fractionsMDH1 activity in serially titrated HeLa cell fractions (Cyto = Cytoplasmic, Mito = Mitochondrial and Nuc = Nuclear) demonstrates the assay’s specificity to MDH1.
