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Senescence-associated beta-galactosidase, along with p16 Ink4A, is regarded to be a biomarker of cellular senescence. [ 1 ] [ 2 ] Its existence was proposed in 1995 by Dimri et al. [ 3 ] following the observation that when beta-galactosidase assays were carried out at pH 6.0, only cells in senescence state develop staining.
β-Galactosidase (EC 3.2.1.23, beta-gal or β-gal; systematic name β-D-galactoside galactohydrolase) is a glycoside hydrolase enzyme that catalyzes hydrolysis of terminal non-reducing β-D-galactose residues in β-D-galactosides. (This enzyme digests many β-Galactosides, not just lactose.
Overview of the use of the SOS response for genotoxicity testing. The SOS chromotest is a biological assay to assess the genotoxic potential of chemical compounds. The test is a colorimetric assay which measures the expression of genes induced by genotoxic agents in Escherichia coli, by means of a fusion with the structural gene for β-galactosidase.
When the target gene is not found in the vector, the alpha fragment gene would be active, producing the alpha fragment and allowing for B-galactosidase to gain its activity. To trace the activity of B-galactosidase a colorless analog of lactose is used, X-gal. The hydrolysis of X-gal by B-galactosidase produces galactose, a blue colored compound.
The presence of an active β-galactosidase can be detected by X-gal, a colourless analog of lactose that may be cleaved by β-galactosidase to form 5-bromo-4-chloro-indoxyl, which then spontaneously dimerizes and oxidizes to form a bright blue insoluble pigment 5,5'-dibromo-4,4'-dichloro-indigo. This results in a characteristic blue colour in ...
ortho-Nitrophenyl-β-galactoside (ONPG) is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity. [1] This compound is normally colorless. However, if β-galactosidase is present, it hydrolyzes the ONPG molecule into galactose and ortho-nitrophen
X-gal itself is colorless, so the presence of blue-colored product may therefore be used as a test for the presence of active β-galactosidase. This also allows for bacterial β-galactosidase (so called lacZ ) to be used as a reporter in various applications. [5] Similarly, Xαgal is used as a reporter compound for α-galactosidase (e.g. Mel1 ...
When the characteristic symptoms of galactosialidosis are suspected, patients can undergo specific testing to confirm their diagnosis. One common method includes enzyme assays which measure the activity of neuraminidase-1 and beta-galactosidase. [4] Decreased levels in enzymatic activity indicate a deficiency in cathepsin A.