PEPCK Assay Kit (Phosphoenolpyruvate Carboxykinase) (ab239714)
Key features and details
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Suspension cells, Tissue
Overview
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Product name
PEPCK Assay Kit (Phosphoenolpyruvate Carboxykinase) -
Detection method
Colorimetric -
Sample type
Tissue, Adherent cells, Suspension cells -
Product overview
PEPCK Assay Kit (Phosphoenolpyruvate Carboxykinase) (ab239714) provides a quick and easy way for measuring PEPCK activity in various samples.
In this assay, Phosphoenolpyruvate Carboxylase is coupled with a set of enzymes that convert PEP and carbonate into a series of intermediates and hydrogen peroxide, which in turn, reacts with a probe and converted generating a colorimetric signal (OD: 570 nm). The color intensity is directly proportional to the amount of active Phosphoenolpyruvate Carboxylase present in samples.
The assay is simple, sensitive, high-throughput adaptable and can detect less than 10 µU of Phosphoenolpyruvate Carboxykinase activity per sample.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests PEPCK Assay Buffer 1 x 25ml PEPCK Converter 1 vial PEPCK Developer 1 vial PEPCK Positive Control 1 vial PEPCK Probe (in DMSO) 1 x 0.2ml PEPCK Substrate Mix 1 vial Pyruvate Standard (100 nmol/µl) 1 x 100µl -
Research areas
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Relevance
Phosphoenolpyruvate Carboxykinase is an enzyme which belongs to the lyase family. In the presence of GTP, it catalyzes the reversible conversion of oxaloacetate (OAA) into phosphoenolpyruvate (PEP), GDP and CO2. In humans, two isoforms of PEPCK are found: cytosolic form (PEPCK-C, also called PCK1) and mitochondrial form (PEPCK-M). PEPCK-C is a rate-controlling step in gluconeogenesis. Recent studies found abnormal concentrations of PEPCK in diabetic mice. Therefore, accurate measurement of Phosphoenolpyruvate Carboxykinase activity is valuable for both, mechanistic and therapeutic studies.
Images
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Pyruvate standard curve.
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Phosphoenolpyruvate Carboxykinase (PEPCK) kinetic activity measured in lysates: HEK-293 cells (66 µg), rat liver (223 µg) and rat kidney (145 µg). -
PEPCK Specific Activity in biological samples.