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).
K5130 01
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The Thermo Fisher Scientific K5130-01 is a lab equipment product. It is a core component used for [core function]. The product specifications and technical details are available from the manufacturer.
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Lab products found in correlation
3 protocols using k5130 01
1
Recombinant DENV2 NS1 Protein Expression
2
Drosophila Expression System for Protein Production
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The mosGCTL genes without signal sequences were cloned into pMT/BiP/V5-His A (V4130-20; Invitrogen) for expression in S2 cells. The primers were shown in Table S3 . The generation of stable cells was described in the product manual of the Drosophila expression system (K5130-01; Invitrogen). The stable cell lines were amplified in regular medium in a 175 cm2 flask and then transferred into spinner flasks with serum-free medium (10486-025; Gibco) for protein expression. The cells were cultured for 3 days and induced with 500 µM copper sulfate for 4 days. The supernatant was centrifuged, filtrated, and then concentrated for purification with a Talon metal affinity resin (635515; Clontech). The protein purity was checked by SDS-PAGE and immunostaining with an anti-V5 mouse monoclonal antibody (M167-3; MBL, Japan).
3
Recombinant DENV2 NS1 Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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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