Authors
Wei Guo and Hong Wu
Summary
Rosa26-LacZ transgenic reporter mice are a very common reporter line to determine Cre activity (1). Since the disruption of target gene(s) and the expression of the Rosa26-LacZ transgene are both controlled by the same Cre recombinase, LacZ expression/activity can serve as a useful marker of the cells with targeted gene disruption. X-gal staining is a common assay used to detect LacZ+ cells in many tissues. In 1988, Nolan, et. al. (2) developed FACS-Gal analysis, a FACS-based detection of LacZ+ cells with a fluorogenic substrate, FDG, of galactosidase (LacZ enzyme). Galactosidase cleaves FDG and releases a fluorescence product FITC (3). However, it is common that LacZ signals varies from sample to sample. Here we establish a procedure to ensure reliable and consistent results for the whole hematopoietic system, with the exception of terminally differentiated erythroid cells, or red blood cells that express much lower LacZ proteins.Further details
The protocol was published on Protocol Exchange in 2008. To see the entire protocol, click on the source link.Advertisement
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