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Alpha Galactosidase (α-Gal) Activity Assay Kit (Fluorometric)

The most sensitive assay to measure α-Gal activity
Catalog #: K407
$440.00

Product Details

Cat # +Size K407-100
Size 100 assays
Kit Summary • Detection method: Fluorescence (Ex/Em = 360/445 nm)
• Application:Measurement of α-Galactosidase activity in various samples
Detection Method Fluorescence (Ex/Em = 360/445 nm)
Species Reactivity Eukaryotes
Applications • Measurement of α-Galactosidase activity in various samples
Features & Benefits • Quick and easy method to measure Alpha Galactosidase activity.
• Detection limit: < 0.1 µU.
• Specific: does not detect beta galactosidase activity
Kit Components • α-Gal Assay Buffer
• α-Gal Stop Solution
• α-Gal Substrate
• 4-Methylumbelliferone Standard
• α-Gal Positive Control
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Alpha-Galactosidase (α-Gal; EC 3.2.1.22) hydrolyzes alpha-galactosyl moieties found in glycolipids and glycoproteins. In mammals, α-Gal hydrolyzes poly- and oligosaccharides commonly found in dietary sources that are difficult to digest. Therefore, α-Gal is used in dietary supplements that help to reduce the production of intestinal gases due to consumption of certain foods. It is known total α-Gal activity is due to two major isozymes with unique, yet different thermostability profiles. Alpha-Galactosidase A, is thermolabile and represents approximately 90% of total α-Gal activity. Fabry Disease, a lysosomal disease disorder, is characterized by mutations in alpha-Galactosidase A. These mutations cause abnormal accumulation of glycosphingolipids in lysosomes. BioVision’s Alpha Galactosidase Activity Assay Kit provides a simple, rapid way to monitor total α-Gal activity in wide variety of biological samples. In this kit, α-Gal cleaves a synthetic specific substrate releasing a fluorophore, which can be easily quantified (Ex/Em= 360/445 nm). The assay is specific, sensitive and can detect as low as 0.1 µU of α-Galactosidase activity.


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Your protocol suggests Sample Preparation for cells as follows: Homogenize pelleted cells with 100 µl ice-cold alpha-Gal Assay Buffer and keep on ice for 10 min. But, it seems like cells were not properly disrupted using alpha-Gal Assay Buffer. Can I use the RIPA (lysis buffer) for homogenizing cells and then dilute with alpha-Gal assay buffer?
We suggest using dounce homogenizer or a probe sonicator (gentle sonication - few short pulses) to break open your cells and keep samples on ice during the process. Just keeping samples in Assay Buffer is not enough to lyse them. In addition to adding the Assay Buffer which has a detergent, we suggest applying some mechanical force to break open the cells.