Protocol SDS

Application

• Hexokinase activity determination in biological samples (e.g. microorganisms, plasma, serum, cultured cells, tissue and culture media.)

Key Features

• Fast and sensitive. Linear detection range (20 μL sample): 0.22 to 100 U/L for 15 min reaction.
• Convenient. Homogeneous “mix-incubate-measure” type assay.
• High-throughput. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.

Method

• OD565nm

Samples

• Biological samples (e.g. microorganisms, plasma, serum, cultured cells, tissue, and culture media.)

Species

• All

Procedures

• 15 min

Size

• 100 tests in 96-well plate

Detection Limit

• 0.22 U/L

Shelf Life

• 6 months

More Details

HEXOKINASE (E.C. 2.7.1.1) is a mitochondrial and cytosolic enzyme and the first committed step in the glycolysis pathway. Hexose sugars, mainly glucose, are phosphorylated using ATP to form glucose-6-phosphate. Hexokinases are implicated in a myriad of human diseases, including cancer and diabetes. BioAssay Systems’ non-radioactive, colorimetric Hexokinase assay involves a stepwise reaction based on the reduction of the tetrazolium salt MTT in an NADPH-coupled enzymatic reaction to a colored product which exhibits an absorption maximum at 565 nm. The increase in absorbance at 565 nm is proportional to the enzyme activity.