Protocol tips
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The RIP1 8-322-His baculovirus (termed RIP1-His) was generated using the BD BaculoGold Transfection Buffer A & B Set (BD Biosciences) according to the manufacturer’s protocol by transfecting 2 μg of pVL1392-RIP1 8-322-His plasmid and 0.5 μg of linearized BaculoGold Bright DNA (BD Biosciences) into 2×106 Sf9 cells. The RIP1-His bacuolvirus was amplified to passage five (P5). Cdc37 baculovirus (FoldHelper-37, AB Vector, LLC) was also amplified in Sf9 cells. |
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| Protocol tips |
| The RIP1 8-322-His baculovirus (termed RIP1-His) was generated using the BD BaculoGold Transfection Buffer A & B Set (BD Biosciences) according to the manufacturer’s protocol by transfecting 2 μg of pVL1392-RIP1 8-322-His plasmid and 0.5 μg of linearized BaculoGold Bright DNA (BD Biosciences) into 2×106 Sf9 cells. The RIP1-His bacuolvirus was amplified to passage five (P5). Cdc37 baculovirus (FoldHelper-37, AB Vector, LLC) was also amplified in Sf9 cells. |
Publication protocol
Spodoptera frugiperda (Sf9) insect cells were grown in T175 flasks at 27 °C in Sf-900 II SFM medium (Gibco/Invitrogen). The RIP1 8-322-His baculovirus (termed RIP1-His) was generated using the BD BaculoGold Transfection Buffer A & B Set (BD Biosciences) according to the manufacturer’s protocol by transfecting 2 μg of pVL1392-RIP1 8-322-His plasmid and 0.5 μg of linearized BaculoGold Bright DNA (BD Biosciences) into 2×106 Sf9 cells. The RIP1-His bacuolvirus was amplified to passage five (P5). Cdc37 baculovirus (FoldHelper-37, AB Vector, LLC) was also amplified in Sf9 cells.
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