Senescence β-Galactosidase Activity Assay Kit (Fluorescence, Plate-Based) #23833
Product Information
Protocol
Product Description
The Senescence β-Galactosidase Activity Assay Kit (Fluorescence, Plate-Based) uses the β-gal Substrate 4-Methylumbelliferyl β-D-galactopyranoside (4-MUG) to detect Senescence-Associated beta-galactosidase (SA-β-gal) activity. Upon binding to β-gal, 4-MUG is hydrolysed to the fluorescent product 4-MU that can be measured at an excitation wavelength of 360 nm and an emission wavelength of 465 nm. Fluorescent intensity correlates with sample β-gal levels. This quantitative assay uses cellular lysates for determination of SA-β-gal activity. Each kit provides enough reagents to perform up to 100 assays in a 96-well plate format.
Background
β-galactosidase (also known as β-gal) is an essential hydrolase enzyme that catalyzes the hydrolysis of galactose-containing carbohydrates into monosaccharides. Substrates of β-galactosides include lactose, various glycoproteins, ganglioside GM1, and lactosylceramides. β-galactosidase is used widely in molecular biology; for example, isolation of recombinant bacteria during molecular cloning utilizes α-complementation of the bacterial β-galactosidase gene (lacZ) in the presence of a β-gal substrate to identify recombinant clones (1). In cell biology, Senescence-Associated beta-galactosidase (SA-β-gal), defined as β-gal activity at pH 6.0, is a widely used marker of replicative senescence. While initially thought to derive from a unique isoform of β-galactosidase expressed specifically in senescent cells (2), SA-β-gal activity was subsequently shown to result from overexpression and accumulation of β-galactosidase in endogenous lysosomes, and is not specifically required for replicative senescence (3).
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