RESUMEN
This chapter outlines the workflow using the ExpiSf™ Expression System designed for high-density infection of suspension ExpiSf9™ cells. The system utilizes a chemically defined, serum-free, protein-free, and animal origin free medium, making it suitable for recombinant protein expression experiments. The ExpiSf™ chemically defined medium allows efficient transfection and baculovirus production directly within the same culture medium. The ExpiSf™ Expression System Starter Kit provides all necessary components, including cells, culture medium, and reagents needed to infect one (1) liter of cell culture. The system's versatility and animal origin free nature make it a valuable tool for various protein expression studies and biotechnological applications.
Asunto(s)
Baculoviridae , Proteínas Recombinantes , Flujo de Trabajo , Animales , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Baculoviridae/genética , Transfección/métodos , Medios de Cultivo/química , Técnicas de Cultivo de Célula/métodos , Línea Celular , Expresión GénicaRESUMEN
This chapter outlines a rapid detection method to determine the virus titer of your baculovirus stock. Staining of cells with fluorescently labeled gp64 antibody allows for flow cytometer-based quantitation of baculovirus-infected insect cells. In this assay, Sf9 cells are infected with tenfold serial dilutions of the test virus stock, and baculovirus titers are calculated based on the ratio of infected to uninfected cells 13 to 18 h after inoculation.
Asunto(s)
Baculoviridae , Citometría de Flujo , Citometría de Flujo/métodos , Baculoviridae/genética , Animales , Células Sf9 , Carga Viral/métodosRESUMEN
This chapter outlines the use of TOPO cloning for streamlined generation of a recombinant plasmid containing your gene of interest for use in the Bac-to-Bac™ Baculovirus Expression System.
