Invitrogen sf9 cells manual

tion (Bac-to-Bac manual, Lifetechnologies). Sf9 cells transfection and protein expres-sion The verified recombinant bacmid was transfected into Sf9 cells using the method as was described by O'Reilly et al. (). Briefly, Purified recombinant bacmid DNA ( ng) and Cellfectin transfection reagent (Lifetechnologies). infection cycle and the changes in cell morphology vary with the insect cell line and strain of baculovirus used. The meta-bolic condition of the culture and growth medium used also can affect the timing of baculovirus infection. The following morphological changes are typical of monolayer Sf9 cells infected with recombinant AcNPV. www.doorway.ru Size: KB.  · Sf9 Cells Transfection and Recombinant Virus Harvesting. The recombinant bacmid (Bac-VGRNb) was transfected according to Bac-to-Bac expression system manual into insect cells (Invitrogen, USA). Briefly, 8×10 5 Sf9 cells were seeded in each well of a 6-well plate and incubated at 27°C for an overnight. Transfection buffer A and B was.

Gibco® Sf9 cells are commonly used to isolate and propagate recombinant baculoviral stocks and to produce recombinant proteins. Gibco® Sf9 cells are adapted to serum-free suspension culture in Gibco® Sf™ II SFM, which saves significant time and expense associated with the adaptation of cultures. cells/mL, determine viable and total cell counts (see procedure). 3. Expand Sf9 cultures by seeding shake flasks at 3 to 5 x viable cells/mL by diluting cells in pre-warmed growth medium. See Subculturing Cells to maintain and subculture Sf9 cells in suspension or adherent culture. Note: We recommend subculturing cells for a minimum of 3. Several insect lines are recommended for protein expression - Sf9, Sf21or High Five cells™, Mimic™Sf9, all from Invitrogen. We have used the Sf9 cells for several years, and found them satisfactory for our purposes, but it would be wise to check the cell line information to determine which line is best for your purposes.

Protocol for Passaging Suspension Cells. Notes on Subculturing Suspension Insect Cells. The SDSs for all Invitrogen products are available at. 3 ກ.ຍ. Sf9 cells (Expression Systems, LLC) could be used as an alternative for Sf9 E. coli strain DH10Bac, Invitrogen, Cat# 6 ມ.ສ. A variety of insect cell lines and Gibco™ cell culture products are This manual provides an overview of the Bac-to-Bac® Baculovirus.