pONE-30A

Protein Expression Eukaryotic cells - S. frugiperda MBP

Experiment
Protein Expression Eukaryotic cells - S. frugiperda MBP
Product
pONE-30A from László Beinrohr, Institute of Enzymology, Research Centre for
Manufacturer
László Beinrohr, Institute of Enzymology, Research Centre for

Protocol tips

Protocol tips
For the pVL based vectors pONE-30A and -31A, Sf9 insect cells were co-transfected with expression plasmid and linearized baculovirus DNA (Oxford Expression Technologies) in a 6-well plate.

Publication protocol

For the pVL based vectors pONE-30A and -31A, Sf9 insect cells were co-transfected with expression plasmid and linearized baculovirus DNA (Oxford Expression Technologies) in a 6-well plate. Baculovirus was amplified in two further rounds (p1 and p2) as a monolayer culture to produce a virus stock ready for infection of the expression cells. Sf9 insect cells at 2×106 cell/mL density were infected with recombinant baculovirus to produce the protein of interest in 2 mL or 500 mL volume in Insect-Xpress medium (Lonza). The infected cells were incubated for 3 days at 27°C with constant shaking at 110 rpm.

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