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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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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.
|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|
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
Amylase activity measured in biological fluids showing activity (mU) per mL of tested sample. Samples were diluted 2 fold.
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.
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).
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