Overview

  • Product namePyruvate Assay Kit
  • Tests
    100 x 1 assay
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay typeQuantitative
  • Sensitivity
    > 1 µM
  • Range
    1 µM - 200 µM
  • Assay time
    0h 40m
  • Product overview

    Abcam's Pyruvate Assay Kit provides a simple, direct and automation-ready procedure for measuring pyruvate concentration in various biological samples such as blood, cells, culture and fermentation media, etc. In the assay, pyruvate is oxidized by pyruvate oxidase via enzyme reactions to generate color (λ= 570 nm) and fluorescence (at Ex/Em = 535/587 nm). Since the color or fluorescence intensity is proportional to pyruvate content, the pyruvate concentration can be accurately measured. The kit detects 1-200 µM pyruvate.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Pyruvate is a central molecule in metabolism through which sugars enter the citric acid cycle. Pyruvate can be converted to carbohydrates during gluconeogenesis or to fatty acids via acetyl CoA. High levels of pyruvate are associated with liver disease and genetic disorders. Pyruvate has also been used to stimulate metabolism leading to loss of body weight.

  • Tested applicationsFunctional Studiesmore details

Properties

  • RelevancePyruvate is a key intersection molecule in the netwrok of metabolic pathways. Pyruvate can be made from glucose through glycolysism converted back to carbohydrates via gluconeogenesis, or to fatty acids or energy via acetyl CoA. It can also be used to construct the amino acid alanine and be converted to ethanol. Glucose is converted to pyruvate which is then converted into acetyl CoA, the main input for the citric acid cycle (TCA). If insufficent oxygen is available, pyruvate breaks down anaerobically, creating lactate in animals and ethanol in plants and microorganisms.

Applications

Our Abpromise guarantee covers the use of ab65342 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 dilution.

Pyruvate Assay Kit images

  • Pyruvate measured fluorometrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Pyruvate measured colourimetrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Pyruvate measured fluorometrically in biological fluids showing quantity (nmol) per mln of tested cells

  • Pyruvate measured fluorometrically in biological fluids showing quantity (nmol) per mL of tested sample

  • Pyruvate measured colourimetrically in biological fluids showing quantity (nmol) per mL of tested sample

  • Pyruvate colorimetric standard curve.

  • Pyruvate fluorometric standard curve.

  • Quantification of pyruvate in human saliva by colorimetric assay. Saliva sample was centrifuged at 10,000 x g for 10 minutes at 4°C. Supernatant (10 µl) was spiked with known amounts of pyruvate standard (6 nmol) and assayed according to protocol. Calculated concentration of pyruvate: 0.08 ± 0.01 mg/dl.

Protocols

References for Pyruvate Assay Kit (ab65342)

ab65342 has not yet been referenced specifically in any publications.

Product Wall

The laboratory responded that EDTA can interfere with the enzymatic reaction in this assay unless it can be diluted out to bare minimal.

I have spoken with the lab regarding the use of this kit in yeast and bacteria. They agree that while we have optimized this assay protocol for samples of mammalian origin, theoretically this assay should work with samples from multiple origins includi...

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You can use plasma samples with this assay. Optimal dilutions have to be determined using a pilot experiment with different volumes.

Uric acid in urine is going to strongly interfere with the assay so besides doing the normal standard curve, the lab recommends to also do standard curve in the presence of X volume of diluted urine (may be 4-15 ul depends). Due to the presence of uric...

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The lab has advised me that you can keep the buffer at 37°C for some time to see if it helps. The 37°C for a short time should not adversely affect the assay; in fact many clients do keep the buffer bottle at this temp water bath while thawing ...

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Thank you for taking the time to complete our questionnaire and providing this further information. I am sorry to hear you have had difficulty obtaining satisfactory results from our D-Lactate Assay Kit (Colorimetric) (ab83429) and our Pyruvate Assay K...

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Thank you for contacting us.

Start with ˜2X106 cells, suspend the cell pellet 500 μl (or ˜4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the...

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Thank you for your email.

We recommend using this kit with 2x106 cells per 500ul of assay buffer. This cell number however is for guidelines only; the number should be optimized as it varies based on cell line type.

One way to prepare...

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Thank you for your enquiry regarding ab65342.

I apologize for my delayed response but was out of the office yesterday.

You can homogenize about 2 x 10^6 cells with 500 µl of the assay buffer, centrifuge it and take the superna...

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The assay kit ab65342 is capable of detecting pyruvate in human plasma. Please note, our products are for "Research Use Only" (RUO), and are not intended for diagnostic use. Also note that due to the presence of LDH in serum and plasma, care must be ta...

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