LacZ staining on cryosections
As already introduced in Section 3.3, the bacterial lacZ gene can be engineered into the targeting construct so that it is activated upon Cre-mediated gene disruption. Consequently, it can then be used to examine the expression of the disrupted gene. This approach was used in our laboratory to monitor the K5-Cre-driven excision of the $1 integrin gene (Brakebusch et al., 2000). The protocol described below uses the $-gal substrate 5-bromo-4-chloroindoxyl-beta-D-galactopyranoside (X-gal) that produces blue precipitates that are insoluble in fixed cryosections.
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