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MAK072

Sigma-Aldrich

Pyruvate Kinase Activity Assay Kit

sufficient for 100 colorimetric or fluorometric tests

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

usage

sufficient for 100 colorimetric or fluorometric tests

detection method

colorimetric
fluorometric

relevant disease(s)

hematological disorder; cancer

storage temp.

−20°C

General description

Pyruvate kinase (PK) is an enzyme that catalyzes the final step of glycolysis, the transfer of a phosphate group from phospho(enol)pyruvate (PEP) to ADP, yielding one molecule of pyruvate and one molecule of ATP. Pyruvate kinase deficiency, due to defects in pyruvate kinase expression or activity, is the second most common cause of hemolytic anemia.

The Pyruvate Kinase Activity Assay Kit provides a simple and direct procedure for measuring pyruvate kinase activity in a variety of biological samples. Pyruvate concentration is determined by a coupled enzyme assay, which results in a colorimetric (570 nm)/ fluorometric (λex = 535/λem = 587 nm) product, proportional to the pyruvate present. One unit of pyruvate kinase is the amount of enzyme that will transfer a phosphate group from PEP to ADP to generate 1.0 μmole of pyruvate per minute at 25 °C.

Application

Pyruvate Kinase Activity Assay Kit has been used to determine the activity of pyruvate kinase in samples.[1][2][3]

Suitability

Suitable for measuring pyruvate kinase activity in various biological samples such as blood, tissues, and culture cells, etc

Principle

The Pyruvate Kinase Activity Assay Kit provides a simple and direct procedure for measuring pyruvate kinase activity in a variety of biological samples. Pyruvate concentration is determined by a coupled enzyme assay, which results in a colorimetric (570 nm)/ fluorometric (λex = 535/λem = 587 nm) product, proportional to the pyruvate present. One unit of pyruvate kinase is the amount of enzyme that will transfer a phosphate group from PEP to ADP to generate 1.0 μmole of pyruvate per minute at 25 °C.

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Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Aquatic Chronic 3 - Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

10 - Combustible liquids


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Therapeutic and Biochemical Effects of Garlic (Allium sativum) on Acrylamide Toxicity in Rabbits: Glycolytic Pathway.
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The M2 isoform of pyruvate kinase (PKM2) is highly expressed in most cancer cells, and has been studied extensively as a driver of oncogenic metabolism. In contrast, the role of PKM2 in nontransformed cells is little studied, and nearly nothing
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Articles

Warburg effect enhances glucose to lactate conversion in tumor cells, regardless of oxygen levels; impacting cancer metabolism since 1924.

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Warburg effect enhances glucose to lactate conversion in tumor cells, regardless of oxygen levels; impacting cancer metabolism since 1924.

Questions

1–3 of 3 Questions  
  1. Is this assay kit compatible to carry out the assay in Baker's yeast?

    1 answer
    1. This kit may be used with variety of biological samples. However, it has not been tested with yeast samples, hence it cannot be guaranteed as suitable. Suitability would need to be validated by the end user.

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  2. What is the optimal cell quantity for conducting an assay with the Pyruvate Kinase Activity Assay Kit, particularly MAK072, and is this kit appropriate for use with red blood cells?

    1 answer
    1. Optimization for the specific sample type is necessary. Typically, the protocol involves using 1 million cells or 10 mg of tissue suspended in 150-200 microliters of assay buffer.

      Helpful?

  3. 请问一下这个是否适用于红细胞PKR检测,并且是否要破裂细胞,同时有酶标仪和九六孔板即可完成相关检测。

    1 answer
    1. Theoretically, the Pyruvate Kinase (PK) Assay Kit should work with isolated red blood cells (RBC). However, RBC has not been purified or tested with this kit, therefore, no specific protocol recommendations can be made.

      The end-user will have to determine the suitability using a reliable way to isolate RBC. Fresh whole blood coagulates which might negatively influence the isolation of RBC. Hence the isolation of RBC as well as the protocol for the PK Assay kit will have to be optimized for the intended application. This kit is labeled as being able to be used with blood as authors in the following citation used this kit for pyruvate kinase in plasma isolated from whole blood.

      Please see the following publication below which may be beneficial for developing the appropriate method.
      Nguyen Ba Vuong, Ha Van Quang, Bui Ngoc Linh Trang, Dao Hong Duong, Nguyen Linh Toan, Hoang Van Tong,
      Association of PKLR gene copy number, expression levels and enzyme activity with 2,3,7,8-TCDD exposure in individuals exposed to Agent Orange/Dioxin in Vietnam,
      Chemosphere, Volume 329, 2023, 138677, ISSN 0045-6535, https://doi.org/10.1016/j.chemosphere.2023.138677.

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