pFastBac1-sccaIL12opt

Protein Expression Eukaryotic cells - Hi5 IL12 canine

Experiment
Protein Expression Eukaryotic cells - Hi5 IL12 canine
Product
pFastBac1-sccaIL12opt from Geraldo Gileno de Sá Oliveira, Centro de Pesquisas Gonçalo Mon
Manufacturer
Geraldo Gileno de Sá Oliveira, Centro de Pesquisas Gonçalo Mon

Protocol tips

Protocol tips
High-five cells (1 × 106/well) cultured as monolayers in serum free medium in 6-well titration plates were infected with the recombinant baculovirus constructs (AcBac-sccaIL-12, AcBac-sccaIL-12opt, AcBacΔCC-sccaIL-12, or AcBacΔCC-sccaIL-12opt) at MOI of 2, 5 or 10. To assess secreted recombinant protein, the cell culture SN were collected at 24, 48 and 72 h post infection, floating cells were spun down at 500×g for 5 min, and the cell-free SN were stored at −20 °C until use. Recombinant canine IL-12 in the cell culture SN was evaluated by dot-blot assay, using mouse anti-histidine C-terminal antibodies (mouse anti-His-C Ab, Invitrogen) as described below.

Publication protocol

High-five cells (1 × 106/well) cultured as monolayers in serum free medium in 6-well titration plates were infected with the recombinant baculovirus constructs (AcBac-sccaIL-12, AcBac-sccaIL-12opt, AcBacΔCC-sccaIL-12, or AcBacΔCC-sccaIL-12opt) at MOI of 2, 5 or 10. To assess secreted recombinant protein, the cell culture SN were collected at 24, 48 and 72 h post infection, floating cells were spun down at 500×g for 5 min, and the cell-free SN were stored at −20 °C until use. Recombinant canine IL-12 in the cell culture SN was evaluated by dot-blot assay, using mouse anti-histidine C-terminal antibodies (mouse anti-His-C Ab, Invitrogen) as described below.

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Papers

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Manufacturer protocol

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