Protocol SDS

Application

• For evaluation of drugs and screening potential inhibitors of α-amylase.

Key Features

• Safe. Non-radioactive assay.
• Fast and convenient. Homogeneous “mix-incubate-measure” type assay. Can be completed in under an hour at room temperature.
• Robust and High-throughput. A Z’-factor of >0.90 was observed in a 384-well format. Can be readily automated to assay thousands of samples per day.

Method

• FP485/520 nm

Samples

• Compounds that affect α-amylase activity

Species

• All

Procedures

• 40 min

Size

• 400 tests in 384-well plate

Detection Limit

• NA

Shelf Life

• 6 months

More Details

AMYLASE belongs to the family of glycoside hydrolase enzymes that break down starch into glucose molecules by acting on α-1,4-glycosidic bonds. The α-amylases (EC 3.2.1.1) cleave at random locations on the starch chain, ultimately yielding maltotriose and maltose, glucose and “limit dextrin” from amylose and amylopectin. In mammals, α-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 α-amylase inhibition are highly desirable in Research and Drug Discovery. BioAssay Systems’ QuantiFluo™ α-Amylase Inhibitor Screening Kit utilizes fluorescence polarization (FP) to screen for potential α-amylase inhibitors. In this assay, α-amylase cleaves a fluorescent amylose substrate. The decrease in FP is directly proportional to the α-amylase activity in the sample. Inhibition is therefore determined by the increase in FP (λex/em = 485/520 nm).