Assay Genie's alpha-amylase assay method involves two steps: (1). alpha-amylase in the sample hydrolyzes starch and the product is rapidly converted to glucose by alpha-glucosidase and hydrogen peroxide by glucose oxidase; (2). hydrogen peroxide concentration is determined with a colorimetric reagent.
For quantitative determination of alpha-amylase activity
- Sensitive and accurate. Linear detection range 0.3 to 50 U/L alpha-amylase in 96-well plate assay.
- Convenient. The procedure involves adding a single working reagent, incubation for 15 min, followed by the detection reagent and a 20-min incubation and reading the optical density at 585 nm.
|Kit Includes||Assay Buffer (pH 7.0): 20 mL Substrate: 120 mL Detection Reagent: 20 mL Enzyme A: 120 mL Glucose Standard: 1 mL Enzyme B: 120 mL|
|Kit Requires||Pipeting devices, centrifuge tubes, clear flat-bottom 96-well plates, plate reader, and optionally membrane filters.|
|Method of Detection||OD585nm|
|Detection Limit||0.3 U/L|
|Samples||Blood, saliva, urine, agriculture etc|
|Protocol Length||40 min|
|Storage||Store Detection Reagent at 4°C and others at -20°C|
|Shelf Life||6 months|
belongs to the family of glycoside hydrolase enzymes that break down starch into glucose molecules by acting on alpha-1,4-glycosidic bonds. The alpha-amylases (EC 220.127.116.11) cleave at random locations on the starch chain, ultimately yielding maltotriose and maltose, glucose and "limit dextrin" from amylose and amylopectin. In mammals, alpha-amylase is a major digestive enzyme. Increased enzyme levels in humans are associated with salivary trauma, mumps due to inflammation of the salivary glands, pancreatitis and renal failure. Simple, direct and automation-ready procedures for measuring amylase activity are very desirable.