Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay试剂盒(ab109902)

概述

  • 产品名称Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay试剂盒
    参阅全部 Pyruvate dehydrogenase 试剂盒
  • 检测方法Colorimetric
  • 测试
    1 x 96 well plate
  • 样品类型
    Cell culture extracts, Tissue
  • 检测类型Enzyme activity
  • 实验步骤Multiple steps standard assay
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
  • 产品概述

    Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902) can be 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.

  • 说明

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

    Store 5X Stabilizer at -20°C.

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

  • 经测试应用适用于: Functional Studiesmore details
  • 平台Microplate

性能

  • 别名
    • PDC
    • PDH
    • PDHC

应用

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.

应用 Ab评论 说明
Functional Studies Use at an assay dependent dilution.

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit 图像

  • 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)

实验方案

Pyruvate dehydrogenase (PDH) Enzyme Activity Microplate Assay Kit (ab109902)参考文献

This product has been referenced in:
  • Zhang Y  et al. Induction of Posttranslational Modifications of Mitochondrial Proteins by ATP Contributes to Negative Regulation of Mitochondrial Function. PLoS One 11:e0150454 (2016). Functional Studies ; Mouse . Read more (PubMed: 26930489) »
  • Golias T  et al. Hypoxic repression of pyruvate dehydrogenase activity is necessary for metabolic reprogramming and growth of model tumours. Sci Rep 6:31146 (2016). Read more (PubMed: 27498883) »

See all 20 Publications for this product

Product Wall

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 ex...

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It is not necessary to avoid EDTA in the protease inhibitor cocktail. EDTA should not affect the pyruvate dehydrogenase enzyme in the sample or the functioning of the assay.

Thank you for your reply.

The laboratory have advised that probably the best in this situation would be to maintain the DCA in all the cell washes after treatment and during the assay itself without NaF, so that full activation of PDH can be o...

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I contacted the laboratories who have confirmed that for phosphatase inhibition we recommend 10mM NaF for use with this kit. A 500mM stock solution can be prepared in water and stored at -20C.

ab65621 is an appropriate protease inhibitor...

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Please see below for further clarification regarding the following questions:

Is it OK to extract mitos from frozen tissues?

A: yes, you can extract mitos from frozen tissues but be aware that freezen/ thaw cycle would weaken the cell...

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Yes, you can use frozen tissues as sample for all three kits. As a reminder, please follow each individual protocol included with each kit to prepare samples for that particular assay.

Thank you for contacting us.



Nome of the reagents in this kit contain thymine.





I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Read More

Please find the answers to your questions from my colleagues at Miosciences, who are designing this kits:
1. The dephosphorylation modifications can be done after the immunocapture of the PDH enzyme to the plate. Many related concerns and question...

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Product ab110338 contains whole mitochondria in iso-osmotic buffer. There is no need to measure protein concentration of this product; the product is supplied at 5.5 mg/mL. Simply adjust the protein concentration with PBS to 5.3 mg/mL and add dete...

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There is no need to homogenize mitochondrial preparation in PBS.

I'd recommend measuring protein concentration of mitochondrial PBS suspension on an aliquot diluted in a detergent (for example 100x dilution of mitochondria suspension in 0.25%...

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