Hexokinase Activity Assay Kit (Colorimetric) (ab136957)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Adherent cells, Serum, Suspension cells, Tissue, Tissue Extracts
- Sensitivity: 0.1 mU/well
Overview
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Product name
Hexokinase Activity Assay Kit (Colorimetric)
See all Hexokinase kits -
Detection method
Colorimetric -
Sample type
Serum, Tissue, Adherent cells, Suspension cells, Tissue Extracts -
Assay type
Enzyme activity (quantitative) -
Sensitivity
Assay time
1h 00mSpecies reactivity
Reacts with: Mammals, Other speciesProduct overview
Hexokinase Activity Assay Kit (ab136957) uses a simple, sensitive and rapid procedure which can detect hexokinase activity to below 0.1 mU/well.
In the hexokinase assay protocol, glucose is converted to glucose-6-phosphate by hexokinase. The glucose-6-phosphate is then oxidized by glucose-6-phosphate dehydrogenase to form NADH. The NADH reduces a colorless probe to a colored product with strong absorbance at 450 nm.
Hexokinase activity assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 20-60 min
- analyze repeatedly with a microplate reader during the incubationNotes
Hexokinases (HK) are found in many organisms including bacteria, plants and mammals and play an important role in glucose metabolism. Hexokinases phosphorylate glucose and generate glucose-6-phosphate for glycolysis. Hexokinases have four isoforms (HK-I, II, III and IV). HK-I, HK-II and HK-III have low Km, while HK-IV has 100 fold high Km.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.Platform
Microplate readerProperties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests Assay Buffer WM 1 x 25ml Developer (Lyophilized) Red 1 vial Hexokinase Coenzyme Purple 1 vial Hexokinase Enzyme Mix Green 1 vial Hexokinase Substrate White 1 x 1ml NADH Standard ( lyophilized) Yellow 1 vial Positive Control Orange 1 vial -
Research areas
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests Assay Buffer WM 1 x 25ml Developer (Lyophilized) Red 1 vial Hexokinase Coenzyme Purple 1 vial Hexokinase Enzyme Mix Green 1 vial Hexokinase Substrate White 1 x 1ml NADH Standard ( lyophilized) Yellow 1 vial Positive Control Orange 1 vial -
Research areas
Images
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Functional Studies - Hexokinase Assay Kit (Colorimetric) (ab136957)
Hexokinase Activity measured in cell lysates showing quantity (nmol) per 1 mln cells after 35 min of incubation.
Samples with the concentration of 5e6 cells/mL were used. Samples were diluted 1-27 fold.
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Functional Studies - Hexokinase Assay Kit (Colorimetric) (ab136957)Hexokinase Activity measured in tissue lysates showing quantity (nmol) per mg of extracted protein after 35 min of incubation.
Protein concentration for samples varied from 13 mg/mL to 43 mg/mL. Samples were diluted 1-9 fold.
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Functional Studies - Hexokinase Assay Kit (Colorimetric) (ab136957)Standard curve: mean of duplicates (+/- SD) with background reads subtracted -
Example dataNADH standard curve (a). Hexokinase activity in Positive Control and Jurkat cell lysate (40 µg) (b). -
Assay ProcedureIn this kit glucose is converted to glucose-6-phosphate by hexokinase; the glucose-6-phosphate is oxidized by glucose-6-phosphate dehydrogenase to form NADH, which reduces a colorless probe to a colored product with strong absorbance at 450 nm.
