Total protein was extracted from cells in RIPA Lysis buffer with protease and phosphatase inhibitor cocktail. Total protein amount was quantified by Bradford assay. 15μg of protein was subjected to electrophoresis using NuPAGE Novex Cel System (Life Technologies, Carlsbad, CA) and transferred to nitrocellulose membrane using iBlot Gel Transfer System (LifeTechnologies, Carlsbad, CA) according to manufacturer’s instructions. Anti-hypocretin receptor-1 antibody (ab68718, Abcam, Cambridge, MA) and anti-βactin antibody (clone 13E5, Cell Signaling, Danvers, MA) were used.
Nupage novex cel system
Manufactured by Thermo Fisher Scientific
The NuPAGE Novex Gel System is a laboratory equipment used for protein separation and analysis. It provides a platform for electrophoretic separation of proteins in polyacrylamide gels. The system includes components necessary for sample preparation, gel casting, and electrophoresis. It is designed to offer consistent and reliable protein separation performance.
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2 protocols using nupage novex cel system
1
Western Blot Analysis of Hypocretin Receptor-1
2
Western Blot Analysis of Hypocretin Receptor-1
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Total protein was extracted from cells in RIPA Lysis buffer with protease and phosphatase inhibitor cocktail. Total protein amount was quantified by Bradford assay. 15μg of protein was subjected to electrophoresis using NuPAGE Novex Cel System (Life Technologies, Carlsbad, CA) and transferred to nitrocellulose membrane using iBlot Gel Transfer System (LifeTechnologies, Carlsbad, CA) according to manufacturer’s instructions. Anti-hypocretin receptor-1 antibody (ab68718, Abcam, Cambridge, MA) and anti-βactin antibody (clone 13E5, Cell Signaling, Danvers, MA) were used.
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