Hexokinase Activity Assay
Assay Genie's Hexokinase Activity Assay kit provides a quick and easy method for monitoring Hexokinase (HK) activity in a wide variety of samples. In this assay, HK converts glucose into glucose-6-Phosphate, which in turn undergoes a series of reactions and reduce to generate intense fluorescent product (Ex/Em = 535/587 nm). The assay is simple, sensitive and high-throughput adaptable and can detect as low as 2 µU of HK activity.
Hexokinase Activity Assay Kit Data
|Product Name:||Hexokinase Activity Assay|
|Detection Method:||Fluorometric (Ex/Em = 535/587 nm)|
|Sample Types:|| |
|Storage:||Store the kit at -20°C, protect from light|
|Shipping Conditions:||Gel Pack|
|Usage:||For research use only. Not for use in humans.|
Hexokinase Activity Assay Kit Components
|HK Assay Buffer||25 ml|
|GenieProbe (in DMSO)||0.4 ml|
|HK Substrate||1 ml|
|HK Enzyme||1 vial|
|HK Developer||1 vial|
|HK Positive Control||1 vial|
|NADPH Standard (200 nmol)||1 vial|
Unit Definition: One unit of Hexokinase is the amount of anzyme that egnerates 1.0µmol of NADPH per min at pH 8.0 at 250C.
Figure: (a)NADPH Standard Curve (b) Kinetic measurement of HK Specific Activity in lysates prepared from orchid leaf (4µg) MCF-7 (0.56µg), Jurkat (0.65µg) and HeLa (0.62µg) cells. (c) HK Specific Activity of samples mentions in fig. b. Assays were preformed following Kit Protocol.
Hexokinase (EC 188.8.131.52; also called HK, 6-Phosphate Glucose Kinase, ATP: D-Hexose 6-Phosphotransferase, ATP-dependent Hexokinase) is found in many organisms including bacteria, plants and mammals. Hexokinases play an important role in glucose metabolism. There are four isoforms (HK-I, II, III and IV). HK-I, HK-II, and HK-III have low Km, while HK-IV (also called Glucokinase) has 100-fold high Km for glucose. Hexokinase deficiency leads to diseases such as X-linked muscular dystrophy and rare autosomal recessive hemolytic anemia. On the other hand, increased hexokinase activity is detected in various human tumors and is associated with metastasis.