pFastBac1- B/Brisbane/60/2008-NP

Protocol tips

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	To produce baculovirus for NP expression, recombinant bacmids were mixed with cellfectin reagent in Sf-900 SFM without supplements or antibiotics according to the manufacturer’s instructions. The transfected cells were then incubated at 27°C for 72 h with daily monitoring for cytopathic effect (CPE). Baculoviruses directing expression of NP from influenza A/reassortant/NYMC X-179 or B/Brisbane/60/2008 were harvested from Sf9 cell culture supernatant at 72 h posttransfection.

Protocol tips

To produce baculovirus for NP expression, recombinant bacmids were mixed with cellfectin reagent in Sf-900 SFM without supplements or antibiotics according to the manufacturer’s instructions. The transfected cells were then incubated at 27°C for 72 h with daily monitoring for cytopathic effect (CPE). Baculoviruses directing expression of NP from influenza A/reassortant/NYMC X-179 or B/Brisbane/60/2008 were harvested from Sf9 cell culture supernatant at 72 h posttransfection.

Publication protocol

"Sf9 insect cells from Spodoptera frugiperda were cultured in suspension at 27°C in Sf-9 00 serum free medium (SFM; Thermo Fisher Scientific, USA) supplemented with 1% penicillin/streptomycin (Sigma-Aldrich, USA). To produce baculovirus for NP expression, recombinant bacmids were mixed with cellfectin
reagent in Sf-900 SFM without supplements or antibiotics according to the manufacturer’s instructions. The transfected cells were then
incubated at 27°C for 72 h with daily monitoring for cytopathic effect (CPE). Baculoviruses directing expression of NP from influenza A/reassortant/NYMC X-179 or B/Brisbane/60/2008 were harvested from Sf9 cell culture supernatant at 72 h posttransfection. Collected supernatant was cleared by centrifuging at 3,000 rpm for 5 min and transferred to fresh 15-ml tubes as the P1 viral stock. The P1 virus was amplified to generate high-titer P3 virus. Viable cell-density, cell viability, and cell size were determined for Sf9 cells infected with P3 viruses at viral dilutions of 1:100, 1:1000, and 1:10,000, or mock infected."

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Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - S. frugiperda Influenza NP using pFastBac1- B/Brisbane/60/2008-NP from Moo-Seung Lee, Department of Biomolecular Science, KRIBB School .

Manufacturer protocol

Download the product protocol from Moo-Seung Lee, Department of Biomolecular Science, KRIBB School for pFastBac1- B/Brisbane/60/2008-NP below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - S. frugiperda Influenza NP using pFastBac1- B/Brisbane/60/2008-NP from Moo-Seung Lee, Department of Biomolecular Science, KRIBB School . Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.