Hexokinase II Inhibitor Assay Kit (Colorimetric) (ab211114)
Key features and details
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 30 min
Overview
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Product name
Hexokinase II Inhibitor Assay Kit (Colorimetric)
See all Hexokinase II kits -
Detection method
Colorimetric -
Assay time
0h 30m -
Product overview
Hexokinase II Inhibitor Screening Kit (Colorimetric) (ab211114) provides a sensitive, quick, and easy method for screening potential inhibitors of Hexokinase II (HK-II) in less than 30 minutes. The assay is based in the ability of HK-II to convert glucose into glucose-6-phosphate. Glucose-6-phosphate is oxidized by glucose-6- phosphate dehydrogenase to form NADH, which reduces a probe that shows strong absorbance at OD 450 nm. In the presence of a HK-II inhibitor such as Bromopyruvic Acid, the reaction is impeded, decreasing the rate or extent of generation of HK-II–dependent absorbance at OD 450 nm.
This simple and high-throughput adaptable assay kit can be used to screen, study or characterize potential inhibitors of Hexokinase II.
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Notes
Hexokinases (HK, EC: 2.7.1.1) are found in many organisms including bacteria, plants and mammals and play an important role in glucose metabolism. The Hexokinase family phosphorylates glucose and generates glucose-6-phosphate for glycolysis. Four Hexokinase isoforms (HK-I, II, III and IV) are found in numerous species. HK-I, HK-II and HK-III have low affinity for glucose, one of their natural substrates, while HK-IV show high affinity for this monosaccharide. Recent studies have found increased hexokinase activity in various human metastatic tumors. Moreover, Hexokinase II (HKII) is the main isoform of the Hexokinases and is responsible for malignant phenotypes. HKII binds to the outer mitochondrial membrane via the Voltage-Dependent Anion Channel (VDAC), a Porin-like protein. HKII has become an attractive therapeutic target for its role in cancer metastasis.
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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 Hexokinase II (Human) 1 x 10µl HK Assay Buffer 1 x 25ml HK Coenzyme (25 mg) 1 vial HK Converter (10 U) 1 vial HK Developer (30 mg) 1 vial HK Inhibitor Control (Bromopyruvic Acid) (50 µmol) 1 vial HK Substrate 1 x 1ml -
Research areas
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Tissue specificity
Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle. -
Pathway
Carbohydrate metabolism; hexose metabolism. -
Sequence similarities
Belongs to the hexokinase family.
Contains 2 hexokinase domains. -
Domain
The N- and C-terminal halves of this hexokinase show extensive sequence similarity to each other. The catalytic activity is associated with the C-terminus while regulatory function is associated with the N-terminus. Each domain can bind a single glucose and Gluc-6-P molecule. -
Cellular localization
Mitochondrion outer membrane. Its hydrophobic N-terminal sequence may be involved in membrane binding. - Information by UniProt
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Alternative names
- DKFZp686M1669
- Hexokinase 2
- Hexokinase 2 muscle
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Images
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Hexokinase II Inhibitor Screening Kit (Colorimetric) (ab211114)
Typical inhibition curve of human Hexokinase II activity by the hexokinase inhibitor Bromopyruvic Acid. IC50 was determined to be 3 µM. Assay was performed following the kit protocol.
