Amylase Assay Kit (Colorimetric) (ab102523)
- Datasheet
- Specific References (4)
- Protocols
Overview
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Product nameAmylase Assay Kit (Colorimetric)
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Detection methodColorimetric
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Sample typeCell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media
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Assay typeQuantitative
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Sensitivity> 0.2 mU/well
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Assay time0h 40m
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Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Mammals
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Product overview
Amylase Assay kit (Colorimetric) (ab102523) detects activity of a-amylase through a two-step reaction. a-Amylase will cleave the substrate ethylidene-pNP-G7 to produce smaller fragments that are eventually modified by a-glucosidase, causing the release of a chromophore that can then be measured at OD = 405 nm. The assay can detect a-amylase content as low as 0.2 mU.
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Notes
Amylases are enzymes that break starch down to sugar molecules. a-amylase is the major form of amylase found in humans and other mammals as well as an enzyme present in seeds, or in fungi (baker's yeast for instance). a-amylase is a calcium metalloenzyme, completely unable to function in the absence of calcium. In human physiology, both the salivary and pancreatic amylases are major digestive enzymes. Increased enzyme levels in humans are associated with salivary trauma; mumps due to inflammation of the salivary glands, pancreatitis and renal failure. A simple, direct and automation-ready procedure for measuring a-amylase activity is, therefore, very desirable.
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PlatformMicroplate
Properties
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Storage instructionsStore at -20°C. Please refer to protocols.
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Components Identifier 100 tests Amylase Assay Buffer NM 1 x 55ml Amylase Positive Control Red 1 vial Amylase Substrate Mix NM 1 x 5ml Nitrophenol standard Yellow 1 x 150µl -
Research areas
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RelevanceAmylases are enzymes that break starch down to sugar molecules. Alpha-amylase is the major form of amylase found in humans and other mammals as well as an enzyme present in seeds, or in fungi (baker’s yeast for instance). Alpha-amylase is a calcium metalloenzyme, completely unable to function in the absence of calcium. In human physiology, both the salivary and pancreatic amylases are major digestive enzymes. Increased enzyme levels in humans are associated with salivary trauma; mumps due to inflammation of the salivary glands, pancreatitis and renal failure. A simple, direct and automation-ready procedure for measuring alpha-amylase activity is, therefore, very desirable.
Images
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Functional Studies - Amylase Assay Kit (ab102523)Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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Functional Studies - Amylase Assay Kit (ab102523)Amylase activity measured in biological fluids showing activity (mU) per mL of tested sample. Samples were diluted 2 fold.
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Functional Studies - Amylase Assay Kit (ab102523)Amylase activity measured in tissue lysates showing activity (mU) per mg of extracted protein.
Protein concentration for samples varied from 16 mg/mL to 50 mg/mL. Samples were diluted 2 fold.
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Functional Studies - Amylase Assay Kit (ab102523)Sample Test -
Functional Studies - Amylase Assay Kit (ab102523)Image from Mondragon A et al., PLoS One. 2014;9(8):e104873. Fig 3.; doi: 10.1371/journal.pone.0104873.Plasma amylase levels were measured (using ab102523) after 75 days treatment with saline, liraglutide, exendin-4 or sitagliptin. ND, normal chow diet; HFD, high fat diet. p≤0.05, *; p≤0.01, **, n = 3–7 mice.
There were no statistically significant changes in plasma amylase activity in mice that were administered liraglutide or exendin-4 vs mice administered saline. However, administration of sitagliptin to animals on normal diet led to a 1.4-fold increase in amylase activity (p≤0.01, n = 3 per group) and a 1.3- fold increase in mice on a high fat diet (p≤0.01, n = 4 per group).
Protocols
References
This product has been referenced in:
- Merle NS et al. Characterization of Renal Injury and Inflammation in an Experimental Model of Intravascular Hemolysis. Front Immunol 9:179 (2018). Read more (PubMed: 29545789) »
- O'Harte FP et al. Dogfish glucagon analogues counter hyperglycaemia and enhance both insulin secretion and action in diet-induced obese diabetic mice. Diabetes Obes Metab 18:1013-24 (2016). Read more (PubMed: 27357054) »
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