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MDH2 Human Activity Assay Kit (ab119693)

MSCatalog No. MS986

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Overview

Product name

MDH2 Human Activity Assay Kit

Precision

Intra-assay
Sample n Mean SD CV%
1 3 4.1%
Inter-assay
Sample n Mean SD CV%
1 3 13.9%

Tests

1 x 96 well plate

Sample type

Cell culture extracts, Tissue Extracts

Assay type

Enzyme activity

Sensitivity

0.78 µg/ml

Range

0.78 µg/ml - 200 µg/ml

Cross reactivity

Reacts with

Mouse, Rat, Human

Notes

Mitochondrial malate dehydrogenase (MDH2, P40926) is a 35.5 kDa enzyme that catalyzes the conversion of malate into oxaloacetate (using NAD+) and vice versa. (EC 1.1.1.37) Several isozymes of malate dehydrogenase exist, depending on where they are localized in the cell and their specific dependence on NAD+ or NADP+ (only in chloroplasts). There are two main isoforms in eukaryotic cells. One is found in the mitochondrial matrix (MDH2), participating as a key enzyme in the citric acid cycle that catalyzes the oxidation of malate. The other is found in the cytoplasm (MDH1), assisting the malate-aspartate shuttle with exchanging reducing equivalents so that malate can pass through the mitochondrial membrane to be transformed into oxaloacetate for further cellular processes. Because malate dehydrogenase is closely tied to the citric acid cycle, regulation is highly dependent on TCA products. High malate concentrations stimulate MDH activity, and, in a converse manner, high oxaloacetate concentrations inhibit the enzyme. Enzyme activity is enhanced by acetylation.

Storage: All components are shipped cold. Reagent dye, coupler, malate and NAD+ are shipped lyophilized. Before use rehydrate by adding 0.25 mL pure H2O to each tube and vortex each tube thoroughly to dissolve. After hydration unused amounts of these four materials should be stored at -80°C for 6 months. Store all other components at 4°C. This kit is stable for 6 months from receipt.

Properties

Storage instructions

Please see Notes section

Components Identifier 1 x 96 tests
100X CouplerP/N 82060151 x 0.168mg
100X NAD+ P/N 82060131 x 17mg
100X Reagent DyeP/N 82060141 x 16mg
100X Sodium MalateP/N 82095221 x 25mg
10X Blocking BufferP/N 82098021 x 6ml
20X BufferP/N 82030131 x 20ml
Base BufferP/N 82095211 x 24ml
Extraction BufferP/N 82010931 x 15ml
MDH2 MicroplateP/N 82095231 unit
  • ELISA - MDH2 Human Activity Assay Kit (ab119693)ELISA - MDH2 Human Activity Assay Kit (ab119693) image (enlarge)

  • ELISA - MDH2 Human Activity Assay Kit (ab119693)ELISA - MDH2 Human Activity Assay Kit (ab119693) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - MDH2 Human Activity Assay Kit (ab119693)Immunocytochemistry/ Immunofluorescence - MDH2 Human Activity Assay Kit (ab119693) image (enlarge)

Sequence similarities

Belongs to the LDH/MDH superfamily. MDH type 1 family.

Post-translational
modifications

Acetylation is enhanced by up to 67% after treatment either with trichostin A (TSA) or with nicotinamide (NAM) with the appearance of tri-and tetraacetylations. Glucose also increases acetylation by about 60%.

Cellular localization

Mitochondrion matrix.

Target information above from: UniProt accessionP40926 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • mitochondrial
  • Malate dehydrogenase
  • mdh2
  • MDHM_HUMAN

MDH2 Human Activity Assay Kit images:

  ELISA - MDH2 Human Activity Assay Kit (ab119693)

ELISA - MDH2 Human Activity Assay Kit (ab119693)

Figure 1. MDH2 activity measurements of serially diluted cultured HepG2 cell extracts.

  ELISA - MDH2 Human Activity Assay Kit (ab119693)

ELISA - MDH2 Human Activity Assay Kit (ab119693)

Figure 2. MDH2 activity measurements of serially diluted human liver homogenate, rat heart homogenate, and mouse liver homogenate.

  Immunocytochemistry/ Immunofluorescence - MDH2 Human Activity Assay Kit (ab119693)

Immunocytochemistry/ Immunofluorescence - MDH2 Human Activity Assay Kit (ab119693)

Figure 4. MDH2 antibody showing reactivity in a mitochondrial intracellular pattern with immunofluorescence microscopy.

  ELISA - MDH2 Human Activity Assay Kit (ab119693)

ELISA - MDH2 Human Activity Assay Kit (ab119693)

Figures 7. The isoform specificity of the malate activity measured by this kit is confirmed by measuring the MDH activity from different cell fractions. Activity was only detected from the mitochondrial fraction (MDH2), not the cytosol fraction (MDH1).

References for MDH2 Human Activity Assay Kit (ab119693)

ab119693 has not yet been referenced specifically in any publications.

Publishing research using ab119693? Please let us know so that we can cite the reference in this datasheet

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