Beta Galactosidase And X Gal

The gene of β galactosidase from e.

Beta galactosidase and x gal. X gal when cleaved by β galactosidase yields galactose and 5 bromo 4 chloro 3 hydroxyindole 1 the latter then spontaneously dimerizes and is oxidized into 5 5 dibromo 4 4 dichloro indigo 2 an intensely blue product which is insoluble. Beta galactosidase beta gal staining is widely used to demonstrate specific gene expression during evaluation of gene targets in vivo. Although x gal is well known reagent to detect β galactosidase in cell or tissue samples the assay using these reagents require to fix cells or tissues due to the poor cell permeability.

This results in a characteristic blue colour in. 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 method is used for the detection of β galactosidase on formalin fixed frozen sections and it is not suitable for formalin fixed paraffin embedded tissue sections.

X gal stock solution 100mg ml in dimethyl formamaide dmf agar. Substrates of different β galactosidases include ganglioside. β galactosidase β gal catalyzes the hydrolysis of lactose.

Normal cells can permanently lose the ability to proliferate when challenged by potentially oncogenic stress a process termed cellular senescence. These protocols are used to screen bacterial plates for beta galactosidase activity using x gal aka blue white screening. Coli is widely used as a reporter gene assay marker.

X gal is an analog of lactose and therefore may be hydrolyzed by the β galactosidase enzyme which cleaves the β glycosidic bond in d lactose. Senescence associated beta galactosidase sa betagal activity detectable at ph 6 0 permits the identification of senescent cells in culture and mamma. X gal staining is a rapid and convenient histochemical technique used to detect reporter gene expression.

Each of these protocols has its advantages and disadvantages so be sure to check which one works best for you. β galactosidase also called lactase beta gal or β gal is a family of glycoside hydrolase enzymes that catalyzes the hydrolysis of β galactosides into monosaccharides through the breaking of a glycosidic bond. X gal staining protocol for beta galactosidase.