Selected article for: "nitrocellulose membrane and total protein"
Author: Jaïs, Philippe H; Decroly, Etienne; Jacquet, Eric; Le Boulch, Marine; Jaïs, Aurélien; Jean-Jean, Olivier; Eaton, Heather; Ponien, Prishila; Verdier, Fréderique; Canard, Bruno; Goncalves, Sergio; Chiron, Stéphane; Le Gall, Maude; Mayeux, Patrick; Shmulevitz, Maya
Title: C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system
  • Document date: 2019_3_18
    • ID: 6nq7y1qe_25
    Snippet: For C3P3 immunoblotting, HEK-293 and CHO-K1 cells were transfected with the pCMV-FLAGx3-NP868R-(G 4 S) 2 -K1ERNAP(R551S) plasmid, which encodes for the FLAGx3 tag fused in frame to the amino-terminal ends of the NP868R-(G 4 S) 2 -K1ERNAP(R551S) C3P3 enzyme. Cells were lysed in 200 l of CLR buffer and lysate was clarified by spinning for 15 s at 12 000 × g at room temperature. Twenty milligrams of total protein were resolved on 4-12% NuPAGE SDS-p.....
    Document: For C3P3 immunoblotting, HEK-293 and CHO-K1 cells were transfected with the pCMV-FLAGx3-NP868R-(G 4 S) 2 -K1ERNAP(R551S) plasmid, which encodes for the FLAGx3 tag fused in frame to the amino-terminal ends of the NP868R-(G 4 S) 2 -K1ERNAP(R551S) C3P3 enzyme. Cells were lysed in 200 l of CLR buffer and lysate was clarified by spinning for 15 s at 12 000 × g at room temperature. Twenty milligrams of total protein were resolved on 4-12% NuPAGE SDS-polyacrylamide gradient gel (Life Technologies, Carlsbad, CA, USA), and subjected to western blotting onto nitrocellulose Hybond membrane (GE Healthcare, Pittsburgh, PA, USA) overnight at +4 • C.

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