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  • A fast and sensitive alternative for β-galactosidase detection in mouse embryos.

A fast and sensitive alternative for β-galactosidase detection in mouse embryos.

Development (Cambridge, England) (2012-11-08)
Sakthi Sundararajan, Maki Wakamiya, Richard R Behringer, Jaime A Rivera-Pérez
ABSTRACT

The bacterial lacZ gene is widely used as a reporter in a myriad of mouse transgenic experiments. β-Galactosidase, encoded by lacZ, is usually detected using X-gal in combination with ferric and ferrous ions. This assay produces a blue indole precipitate that is easy to detect visually. Here, we show that Salmon-gal in combination with tetrazolium salts provides a more sensitive and faster staining reaction than the traditional β-galactosidase assay in mouse embryos. Using a combination of Salmon-gal and tetranitroblue tetrazolium, we were able to visualize the activity of β-galactosidase in embryos at stages when the customary X-gal reaction failed to detect staining. Our studies provide an enhanced alternative for β-galactosidase detection in expression and cell fate studies that use lacZ-based transgenic mouse lines.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
5-Bromo-4-chloro-3-indolyl β-D-galactopyranoside, ≥98%, powder
Sigma-Aldrich
5-Bromo-4-chloro-3-indolyl β-D-galactopyranoside, Vetec, reagent grade, ≥98%
Roche
NBT, 4-Nitro blue tetrazolium chloride, solution