Invitrogen sf9 manual

 · 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. Introduction This manual is included with Sf9, Sf21, and High FiveŽ insect cells and provides general information about the growth and maintenance of insect cell cultures. The Sf9, Sf21, and High FiveŽ cell lines are suitable for use in expressing recombinant proteins with baculovirus and other insect expression systems (e.g. InsectSelectŽ System).File Size: KB.  · We have been using the Cellfectin® Reagent in our lab for quite some time for transfecting Sf9 and High Five™ cells, mainly baculovirus DNA and occasionally the pMIB/V5-His vectors from Invitrogen’s InsectSelect™ system. For these constructs, transfection resulted in high levels of both virus and protein being produced.

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™ III SFM, which saves significant time and expense associated with the adaptation of cultures. The Sf9 insect cell line is a clonal isolate derived from the parental Spodoptera frugiperda cell line IPLB-SfAE1,2, and is a suitable host for expression of recombinant proteins from baculovirus expression systems3,4 (e.g. Invitrogen’s Bac-to-Bac® and Bac-N-Blue™ Expression Systems). The Sf9 cell line exhibits the following general features. This manual provides general information about the growth and maintenance of insect cell cultures. The Sf9, Sf21, and High Five ™ cell lines are suitable for use in expressing recombinant proteins with baculovirusand other insect expression systems (e.g., InsectSelect ™ System). Contents and storage Contents Cat. No. Amount Storage 1.

manual and O'Reilly et al., ). Several companies (e.g., Invitrogen and Clontech) sell PCR primers that will work for most baculovirus vectors. 3 Cell line specific culturing instructions: Sf9 cells. Maintain cells at a density of – x cells/mL. The ProtoArray® Prospector software and manual are available for free to infect Sf9 insect cells for expression of the recombinant protein of interest.