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The Hexokinase Activity Assay Kit (Fluorometric) (ab211103) provides a simple, sensitive and quick method for monitoring hexokinase (HK) activity in cells, serum, and animal or plant tissues. In this assay, HK converts glucose into glucose-6-phosphate, which in turn undergoes a series of reactions and reduces the sensitive probe to generate an intense fluorescent product that can be easily detected at Ex/Em = 535/587 nm.
This assay can detect as low as 2 µU of HK activity.
Hexokinase (HK, 6-Phosphate glucose kinase, ATP:D-Hexose 6- Phosphotransferase, ATP-dependent hexokinase, EC 184.108.40.206) is responsible for phosphorylating hexoses (six-carbon sugars) to form hexose phosphate. Hexokinases play an important role in glucose metabolism, as glucose is the most important substrate of hexokinases. Hexokinases are found in many organisms including bacteria, plants and mammals. In mammals, there are four isoforms (HK-I, II, III and IV). HK-I, HK-II, and HK-III are referred as “low Km” because of their high affinity for glucose (Km < 1mM), while HK-IV (also known as Glucokinase) has a Km for glucose 100-fold higher and can only phosphorylate glucose when the substrate concentration is high enough.
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.
|ATP (50 µmol)||1 vial|
|HK Assay Buffer||1 x 25ml|
|HK Developer (50 U)||1 vial|
|HK Enzyme (100 U)||1 vial|
|HK Positive Control (500 mU)||1 vial|
|HK Substrate||1 x 1ml|
|NADPH Standard (200 nmol)||1 vial|
|PicoProbe (in DMSO)||1 x 400µl|
Typical NADPH standard calibration curve.
Kinetic curves showing Hexokinase activity in positive control (included in kit), lysates from HeLa (0.62 µg), Jurkat (0.65 µg) and MCF-7 cells (0.56 µg), and lysates from orchid leaf (4 µg).
Hexokinase specific activity in lysates from orchid leaf (4 µg), MCF-7 (0.56 µg), Jurkat (0.65 µg) and HeLa cells (0.62 µg).
ab211103 has not yet been referenced specifically in any publications.
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