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Protocol to measure ß-galactosidase in Drosophila extracts using a CPRG assay

The quantification of ß-galactosidase activity is routinely required by laboratories worldwide. We present a cost-effective, highly replicable, simple technique for quantifying ß-galactosidase-specific activity from crude extracts made from whole organisms or dissected tissues or cells. Extracts are...

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Detalles Bibliográficos
Autores principales: Barwell, Taylor, Seroude, Laurent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9667300/
https://www.ncbi.nlm.nih.gov/pubmed/36595888
http://dx.doi.org/10.1016/j.xpro.2022.101843
Descripción
Sumario:The quantification of ß-galactosidase activity is routinely required by laboratories worldwide. We present a cost-effective, highly replicable, simple technique for quantifying ß-galactosidase-specific activity from crude extracts made from whole organisms or dissected tissues or cells. Extracts are prepared and measured without the need for any specialized equipment, and tissue is ground manually by pestle and measured by colorimetric CPRG and Bradford assays. This protocol describes the assay using Drosophila extracts but could be applied to any biological system of interest. For complete details on the use and execution of this protocol, please refer to Seroude et al. (2002),(1) Poirier et al. (2008),(2) and Barwell et al. (2017).(3)