Overview

  • Product nameMalate Dehydrogenase 2 (MDH2) Activity Assay
  • Detection methodColorimetric
  • 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 typeEnzyme activity
  • Sensitivity
    0.78 µg/ml
  • Range
    0.78 µg/ml - 200 µg/ml
  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    Malate Dehydrogenase 2 (MDH2) Activity Assay (ab119693) is used to determine mitochondrial malate dehydrogenase activity (MDH2) in a sample. The enzyme is captured within the wells of the microplate and activity is determined by following the production of NADH in the following MDH2 catalyzed reaction: malate + NAD → oxaloacetic acid + NADH (↑ Absorbance at 450 nm). 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 450 nm. In each well, ab119693 immunocaptures only native MDH2 from the chosen sample; this removes all other enzymes, including MDH1 in cytosol.


    This product allows researchers to focus on TCA cycle, studying isotype-specific malate dehydrogenase (MDH2) activity assay without the necessity of isolating mitochondria.

  • 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.

  • Tested applicationsSuitable for: Functional Studiesmore details
  • PlatformMicroplate

Properties

Applications

Our Abpromise guarantee covers the use of ab119693 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
Functional Studies Use at an assay dependent concentration.

Malate Dehydrogenase 2 (MDH2) Activity Assay images

  • Figure 1. MDH2 activity measurements of serially diluted cultured HepG2 cell extracts.
  • Figure 2. MDH2 activity measurements of serially diluted human liver homogenate, rat heart homogenate, and mouse liver homogenate.
  • Figure 4. MDH2 antibody showing reactivity in a mitochondrial intracellular pattern with immunofluorescence microscopy.
  • 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).

Protocols

References for Malate Dehydrogenase 2 (MDH2) Activity Assay (ab119693)

This product has been referenced in:
  • Kim EY  et al. Acceleration of adipogenic differentiation via acetylation of malate dehydrogenase 2. Biochem Biophys Res Commun 441:77-82 (2013). Read more (PubMed: 24134846) »

See 1 Publication for this product

Product Wall


You are right we use 2F5AF8 as the capture antibody to purify MDH2 from samples. The immunogen of the antibody is purified MDH2, we do not know the epitope of this antibody, but we know the antibody did not bind to the active site, substrate bindi...

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The MDH2 activity assay kit ab119693 is compatible with frozen tissue.

We would recommend dilution of the protein to a working range using 1X incubation buffer. If loading samples with more than 50% detergent, we suggest adding the same amount of detergent to the protein. Please have care when adding a recombinant pro...

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Thank you for contacting us.

The product referred in your email (ab119693) is not intended to measure Malate in your samples, but the enzymatic activity of Malate dehydrogenase.

We do have a different kit in the catalogue, ab83391, ...

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Our suggestion is to purchase 15% Lauryl Maltoside Solution (ab109858) and then produce the following buffer with the following recipe:


25 mM Hepes
100 mM NaCl
1.5% Lauryl Maltoside
pH 7.4

I hope this is helpfu...

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Thank you for your inquiry.

The lab was able to provide the following information:

The answer can be found at the beginning of the booklet: (page 3)
http://www.abcam.com/ps/products/119/ab119693/documents/ab119693%20%20MDH2%20Act...

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Thank you for contacting Abcam yesterday.

Yes, for activity assays the tissue section may be frozen. The tissue homogenate may also be frozen at -80C. What we don’t recommend is to freeze the extract after addition of the provided deter...

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Thank you for your reply. My colleague Cathrin is not in the office today.

Unfortunately, I do also not know if this kit is compatible with a Tecan reader.

I can suggest to ask the manufacturer of the Tecan reader. They should be ab...

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Thank you for contacting us.

I have some good and some bad news for you. the good ones are that it will work in mice, and that the kits may work with Drosophila samples as we as the homology is for both enzymes compared with the human protei...

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Thank you for contacting us.

I have heard back from the lab with the following information:

All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/micro...

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