EnzyChrom™ α-Amylase Assay Kit

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ProtocolSDS

Application

For quantitative determination of α-amylase activity.

Key Features

Sensitive and accurate. Linear detection range 0.3 to 50 U/L α-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.

Method

OD585nm

Samples

Blood, saliva, urine, agriculture etc

Species

Size

100 tests

Detection Limit

0.3 U/L

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 amylase activity are very desirable. BioAssay Systems' EnzyChrom™ a-amylase assay method involves two steps: (1). α-amylase in the sample hydrolyzes starch and the product is rapidly converted to glucose by α-glucosidase and hydrogen peroxide by glucose oxidase; (2). hydrogen peroxide concentration is determined with a colorimetric reagent.

Does glucose interfere with the assay?

The glucose content of the sample will add a significant background and I would recommend removing it from the sample:- For samples known to contain glucose, use a membrane filter (e.g.Microcon YM-10 from Millipore) to remove glucose: load 50 µL sample in a Microcon YM-10 (10 kDa cutoff) and add 500 µL Assay Buffer.- Centrifuge at 14000 rpm for 30 min, check level of sample, ideally the sample level will be less than 50 µL. Add 500 µL Assay Buffer and repeat the centrifugation. Measure final sample volume with a pipetman and calculate dilution factor n = final sample volume/50 µL.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.

Lee, Sang-Bum, et al (2019). Impacts of whey protein on starch digestion in rumen and small intestine of steers. Journal of Animal Science and Technology 61.2: 98-108. Assay: alpha-Amylase in beef steers duodenal fluid.Han, MJ & Lee SH (2015). An efficient bacterial surface display system based on a novel outer membrane anchoring element from the Escherichia coli protein YiaT. FEMS Microbiol. Lett. 362.1: 1-7. Assay: a-Amylase in Bacillus subtilis (bacteria) cells.Han, MJ, and Seung HL (2015) An efficient bacterial surface display system based on a novel outer membrane anchoring element from the Escherichia coli protein YiaT." FEMS Microbiology Letters 362.1: 1-7. Assay: alpha-Amylase in mammal cells. Abu Khadra, KM et al (2014). Antioxidant Profile of Saliva among Young Men Using Mobile Phones. Jordan Journal of Biological Sciences. 7(4):275-280. Assay: alpha-Amylase in human saliva.Lee, KH et al (2014). Identification of proteins involved in the pancreatic exocrine by exogenous ghrelin administration in Sprague-Dawley rats. J. Animal Sci. Techn. 56.1 : 6. Assay: a-Amylase in rat plasma.Lee, KH et al (2014). Identification of proteins involved in the pancreatic exocrine by exogenous ghrelin administration in Sprague-Dawley rats. Journal of Animal Science and Technology. 56:6. Assay: alpha-Amylase in sprague-dawley rats pancreas tissue. Lee, KH et al.(2014). Identification of proteins involved in the pancreatic exocrine by exogenous ghrelin administration in Sprague-Dawley rats. Journal of Animal Science and Technology 56.1: 6. Assay: alpha-Amylase in rat plasma.Lee, Kyung-Hoon, et al.(2014). Identification of proteins involved in the pancreatic exocrine by exogenous ghrelin administration in Sprague-Dawley rats." Journal of Animal Science and Technology 56.1: 6. Assay: alpha-Amylase in mice plasma. Mohannad N et al (2014). Age-related changes in salivary biomarkers. Journal of Dental Sciences. 9(1): 85-90. Assay: alpha-Amylase in human saliva. Mohannad N et al (2014). Age-related changes in salivary biomarkers. Journal of Dental Sciences. 9(1): 85-90. Assay: a-Amylase in human Saliva.Lee, SB et al (2013). Effect of Oral Administration of Intact Casein on Gastrointestinal Hormone Secretion and Pancreatic alpha-Amylase Activity in Korean Native Steer. Asian-Australasian Journal of Animal Sciences (AJAS). 26(5): 654-660. Assay: a-Amylase in steers (cow) pancreas (duodenal fluid).Iqbal S, et al (2009). Feeding barley grain steeped in lactic acid modulates rumen fermentation patterns and increases milk fat content in dairy cows. J Dairy Sci. 92(12):6023-32. Assay: alpha-Amylase in food grain.To find more recent publications, pleaseclick here.

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.Simply send us your samples:- Fast turnaround - Quality data - Low costPlease email or call 1-510-782-9988 x 2 to request assay service.

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