This kit has not been tested specifically for use in measuring intracellular pyruvate in E. coli. However, other researchers have used this kit successfully. Methanol is not an interfering substance. Pelleted cells may be quenched and subsequently flash frozen in methanol to lyse. Repeated flash freezing may be applied to assure complete lysis. The combined collected supernatant may then be analyzed by following the recommended protocol. Please see the link below to review.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/358/918/mak332bul.pdf
MAK332
Pyruvate Assay Kit
sufficient for 100 colorimetric or fluorometric tests
Synonym(s):
Pyruvic Acid Detection Kit
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€371.00
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1 KIT
€371.00
About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Recommended Products
usage
sufficient for 100 colorimetric or fluorometric tests
input
biological sample(s)
application(s)
pharmaceutical
detection method
colorimetric
fluorometric
relevant disease(s)
gastrointestinal diseases; cancer
storage temp.
−20°C
General description
Pyruvate is a key intermediate in cellular metabolic pathways. Pyruvate can be converted to carbohydrates via gluconeogenesis, to fatty acids or energy through acetyl-CoA, to the amino acid alanine and to ethanol. Abnormal levels of pyruvate have been linked to liver diseases and metabolic disorders. Simple, direct and automation-ready procedures for measuring pyruvate concentrations find wide applications in research and drug discovery.
Application
The Pyruvate Assay Kit may be used for:
- Drug Discovery
- Food and Beverage Testing
- Neurological Disease Research
Features and Benefits
Broad Detection Range: Detect Pyruvate levels accurately across a wide range, from 2 µM to 500 µM, using only 10 μL of sample in a 96-well plate setup, ensuring comprehensive analysis with minimal sample requirement.
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 30 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 30 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Suitability
Suitable for pyruvate detection in biological samples and for studying the effects of drugs on pyruvate metabolism.
Principle
The Pyruvate Assay Kit uses a single Working Reagent that combines pyruvate oxidase and hydrogen peroxide determination in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at λem/ex = 585/530nm is directly proportional to pyruvate concentration in the sample. Linear detection range in 96-well plate: 2 to 500 μM (17 μg/dL to 4.4 mg/dL) pyruvate for colorimetric assays and 0.2 to 50 μM for fluorometric assays.
Other Notes
For additional information on our range of Biochemicals, please complete this form.
Storage Class Code
10 - Combustible liquids
Flash Point(F)
188.6 °F
Flash Point(C)
87 °C
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Varun Kamat et al.
eLife, 10 (2021-11-05)
Oxygen (O2) and other dissolved gases such as the gasotransmitters H2S, CO, and NO affect cell metabolism and function. To evaluate effects of dissolved gases on processes in tissue, we developed a fluidics system that controls dissolved gases while simultaneously
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I want to detect E.coli intracellular pyruvate concentration. Therefore, I need to break cells before I detect it. How can I use this kit? I try to use methanol to deal with cells and use freeze-thaw cycle to break. Will methanol have effect to the kit？
1 answer-
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Can I use the samples in Lactate Assay buffer (Lactate assay kit sigma MAK064) for this kit or does this buffer interfere with the reaction ?
1 answer-
The assay buffer for Kit MAK064 has not been tested to determine suitability with the Kit MAK332.
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