The DENV2 NS1 gene was cloned into a pMT/BiP/myc-His A vector (modified from pMT/BiP/V5-His A, V4130-20, Invitrogen) for expression in Drosophila S2 cells. The cloning primers are shown in Supplementary Table 2 . The procedure used to generate stable cells is described in the manual of the Drosophila expression system (K5130-01, Invitrogen). NS1-expressing, stable S2 cells were then amplified in regular Schneider’s Medium in a 175- cm2 flask and transferred into spinner flasks containing Express Five serum-free medium (10486-025, Gibco) for protein expression. The cells were cultured for 3 days and induced with 500 μM copper sulfate for another 4 days. The supernatant was centrifuged, filtered, and then concentrated for purification using a TALON Purification Kit (635515, Clontech). Protein purity was verified by SDS-PAGE and immunostaining with an anti-myc mouse monoclonal antibody (M047-3, MBL, Japan).
V4130 20
Manufactured by Thermo Fisher Scientific
The V4130-20 is a laboratory equipment product from Thermo Fisher Scientific. It is designed for general laboratory use. The core function of the V4130-20 is to provide a controlled environment for various laboratory applications.
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2 protocols using v4130 20
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Recombinant DENV2 NS1 Protein Expression
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Recombinant DENV2 NS1 Protein Expression
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The DENV2 NS1 gene was cloned into a pMT/BiP/myc-His A vector (modified from pMT/BiP/V5-His A, V4130-20, Invitrogen) for expression in Drosophila S2 cells. The cloning primers are shown in Supplementary Table 2 . The procedure used to generate stable cells is described in the manual of the Drosophila expression system (K5130-01, Invitrogen). NS1-expressing, stable S2 cells were then amplified in regular Schneider’s Medium in a 175- cm2 flask and transferred into spinner flasks containing Express Five serum-free medium (10486-025, Gibco) for protein expression. The cells were cultured for 3 days and induced with 500 μM copper sulfate for another 4 days. The supernatant was centrifuged, filtered, and then concentrated for purification using a TALON Purification Kit (635515, Clontech). Protein purity was verified by SDS-PAGE and immunostaining with an anti-myc mouse monoclonal antibody (M047-3, MBL, Japan).
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