QuantiFluo™ α-Amylase Inhibitor Screening Kit

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. The FP assay greatly reduces background matrix interferences. 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 a 384-well plate (200 tests in 96-well plates)

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’ α-Amylase Inhibitor Screening Kit utilizes fluorescence polarization (FP), a highly reliable and robust technique that significantly reduces background matrix interferences, enabling the effective identification of potential α-amylase inhibitors. In this assay, α-amylase cleaves a fluorescent 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).

Does this kit come with α-amylase?

Yes, this kit comes with a purified human salivary α-Amylase, assay buffer, fluorescent substrate and an inhibitor control (acarbose).

What equipment should I use?

A fluorescence polarization-capable plate reader that can read 485/520 nm.

What buffer should I use with this kit?

If more assay buffer is needed, we recommend 20 mM KPi, 50 mM NaCl, pH 7.0.

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.