X-Gal Staining Kit

Protocol tips

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	Add Fixing Buffer to the dish and incubate for 10-15 minutes at room temperature. Add 1X X-Gal staining solution and incubate the cells between 1-18 hours at 37°C in a humidified incubator.

Protocol tips
Add Fixing Buffer to the dish and incubate for 10-15 minutes at room temperature. Add 1X X-Gal staining solution and incubate the cells between 1-18 hours at 37°C in a humidified incubator.

Publication protocol

Transfection of C2C12 cells (5×106) with 10 μg ScaI–linearized pCALL-DECT was performed with the Amaxa nucleofector system (Amaxa GmbH, Cologne, Germany) as previously described (Ozawa and Kobayashi, 2015). Two days after transfection, cells were incubated in medium containing 100 µg/ml G418 for an additional 5–7 days to isolate drug-resistant colonies. G418-resistant clones were analyzed for their ability to differentiate in differentiation medium, before they were expanded and frozen. The clones were transfected with pSR016 using the calcium phosphate precipitation method, and subjected to selection using puromycin (5 μg/ml). Clones that became positive for DECT expression after Cre introduction were used for further studies. β-galactosidase (LacZ) activity was assayed using a kit, X-Gal Staining (Gelantis, San Diego, CA, USA).

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Manufacturer protocol

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