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Amylase Activity Colorimetric Assay Kit

based on 7 citations in multiple journalsAmylase Activity Colorimetric Assay Kit74.2 4
Sensitive Assay, HTS
Catalog #: K711
$305.00

Product Details

Cat # +Size K711-100
Size 100 assays
Detection Method Absorbance (405 nm)
Species Reactivity Mammalian
Applications The assay can detect α-amylase content as low as 0.2 mU.
Features & Benefits • Simple procedure; takes ~ less than 40 minutes
• Fast and convenient
Kit Components • Amylase Assay Buffer
• Amylase Substrate Mix
• Amylase Positive Control
• Nitrophenol standard (2 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Amylases are enzymes that break starch down to sugar molecules. α-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). α-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 α-amylase activity is, therefore, very desirable. BioVision’s α-amylase Assay uses ethylidene-pNP-G7 as the substrate. Once the substrate has been specifically cleaved by α-amylase, the smaller fragments produced can be acted upon by α-glucosidase, which causes the ultimate release of the chromophore that can then be measured at 405 nm. The assay can detect α-amylase content as low as 0.2 mU.


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Can bacterial amylases be detected by this kit?
We have not tested this kit with bacterial samples but bacterial samples could work as long as there is active amylase in them and the sample volume is optimized to get numbers in the linear range of the std. curve. For gram positive bacteria use bacteria specific-reagents to break the cell wall: http://www.biovision.com/ezlys-tm-bacterial-protein-extraction-reagent-3692.htmlWe have not tested this kit with this bacterial reagent.
How long and at what temperature can tissue homogenates be stored for this assay?
For best results we always recommend running the assay as soon as possible with fresh homogenates. If needed the homogenates can be stored at -80C in aliquots to prevent freeze-thaw cycles. 1 week up to 1 month should be fine but long term storage might result in deterioration of sample quality. Enzymes frequently lose activity or are degraded upon storage.
Will this kit work for beta or gamma amylases as well?
This kit is specific for alpha-amylase activity
Is there an upper limit for this kit regarding alpha amylase concentration?
To be able to calculate Amylase activity in samples, it is essential to get values within the linear range of the std curve. The highest datapoint for the std is 20 nmol of Nitrophenol per well. As mentioned on our datasheet, I unit of amylase generates 1.0µmol of nitrophenol per min at pH 7.20 at 25°C. Hence the maximum amount of enzyme activity within the linear range of the std curve would be 20 mU per well. The sample amount needed to get values within the linear range of the std. curve is sample dependent and has to be optimized in a pilot assay by the user.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
Can individual components of this kit be purchased separately?
Yes, any of the kits components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions.
Tilokwattanothai, A. et al., (2017) Hypoglycemic and hypolipidemic properties of herbal tea on Wistar rat, Chiang Mai Veterinary Journal, 2017, 15(1): 25-35
Anuwat T et al., (2017) Hypoglycemic and hypolipidemic properties of herbal tea on Wistar rat, Chiang Mai Veterinary Journal, 2017, 15(1): 25-35
Pluschke A et al., (2017) Protection of α-amylase from proteolysis by adsorption to feed components in vitro and in the porcine small intestine, Animal Prod Sc, 2017, online
Cui K et al., (2017) Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus (Bloch 1790), Aquaculture Res, 2017, 48(8):4533-4544
Bansemer, Matthew S. et al. (2016) Dietary inclusions of dried macroalgae meal in formulated diets improve the growth of greenlip abalone (Haliotis laevigata), J Appl Phycol (2016) 28: 3645.
For more citations of this product click here