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MitoSciences (ab109902)

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

Overview

  • Product namePyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay KitSee all Pyruvate dehydrogenase kits ...
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts, Tissue
  • Assay typeEnzyme activity
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    This rapid activity microplate kit is used to determine the activity of PDH in a human, bovine, mouse, or rat sample. The PDH enzyme is immunocaptured within the wells of the microplate and activity is determined by following the reduction of NAD+ to NADH, coupled to the reduction of a reporter dye to yield a colored reaction product with an increase in absorbance at 450 nm at room temperature. Included for performance of the activity assay are buffer, detergent, reagent mix, and a 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.

    This assay is optimized for use with whole tissue extract when the amount of total material available for assay is 20-100 µg or more. If using cell extract of cultured cells 1 mg of material is required due to the very low levels of enzyme and reduced levels of mitochondria in the extract.

  • Notes

    Store 20X Buffer, Detergent, and Microplate at 4°C.

    Store 5X Stabilizer and 100X Reagent Dye at -20°C.

    Store 20X Reagent Mix and 100X Coupler at -80°C.

  • Tested applicationsFunctional Studies more details

Properties

  • Alternative names
      PDCPDHPDHC

Applications

Our Abpromise guarantee covers the use of ab109902 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Functional Studies FuncS

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit images

  • Mitochondria, tissue extracts and whole cultured cell extracts all show linear relationships between signal and sample load at limiting concentrations. The rates shown were determined as change in OD over time, and these are best represented as change in milliOD per minute.
  • Example of microplate reader recorded data from bovine heart mitochondria (100 µg/well) (top trace) and 2-fold dilutions (stepwise lower traces) using Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
  • Schematic diagram showing the reaction involved in Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902).
  • Principle of Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

Protocols

References for Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)

This product has been referenced in:
  • Ota A  et al. Effects of aripiprazole and clozapine on the treatment of glycolytic carbon in PC12 cells. J Neural Transm : (2012). Read more (PubMed: 22392058) »
  • Thomas SN  et al. Quantitative proteomic analysis of mitochondrial proteins reveals prosurvival mechanisms in the perpetuation of radiation-induced genomic instability. Free Radic Biol Med 53:618-28 (2012). Read more (PubMed: 22569412) »

See all 7 Publications for this product

Product Wall

Displaying 1 - 10 of 62 results for Abreviews and Q&A

Thank you for contacting us. I have heard back from the lab: Yes, liver tissues were tested with ab109902; for example extract of mouse liver homogenate. For extract of liver homogenate, 100 – 500 ug/200 uL should be used, please see attached examp...

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Ich kann bestätigen, dass die Negativkontrolle (nur Puffer) auch einen messbare Optische Dichte hat. Diese OD sollte von allen Proben abgezogen werden: d.h. der Hintergrund wird von allen Proben abgezogen.

Die eigentlichen Proben ...

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To measure the endogenous PDH activity in this case we would recommend to add the protease inhibitors, the ATP-depleting system and also the phosphatase inhibitors already to the sample during the whole tissue homogenization, since mitochondria will be br...

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We advise use of PBS for homogenization of muscle tissue. We have not tested the use of this kit with fat tissue. If you have a favorite homogenization buffer, we would advice to prepare the homogenates of both tissues in both buffers in parallel , run th...

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As a positive control, I would highly recommend MS802 (ab110338). This product contain bovine heart mitochondria. The mitochondria need to be extracted.

Thank you for your response.

There is an instrument called "Dipstick Reader" which has been developed to read the signal. The Protocol Booklet and Measurement Program Instruction Manual can be downloaded through the website:

...

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The total protein concentration of a homogenate should be determined just after the homogenization. No spin is involved. 2. Yes, the homogenate is cloudy. Yes, dilute an aliquot of the homogenate in 0.25 % SDS to determine total protein concentra...

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When using tissue homogenates, to determine concentration you will not need to add detergent. After determining concentration solubilzation will occur by adding detergent in step 2. You will want to spin at RCF(xg) 1000 for 10 minutes at +4C. We do recom...

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Thank you for your enquiry.

We have not tested Arabidopsis with this kit. The immunogen used to generate the capture antibody in this kit was Purified PDH protein from Pig Heart. Therefore, we are unable to predict homology with your species. I...

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Vielen Dank für Ihren Anruf.
Ich habe nun Nachricht von Labor erhalten und kann Ihnen Folgendes ausrichten:
"I would highly recommend storing whole tissue homogenate or whole mitochondria preparations over an extracted sample. In any case, the s...

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