pFastBac1-sccaIL-12

Protocol tips

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	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.

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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Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - Hi5 IL12 canine using pFastBac1-sccaIL-12 from Geraldo Gileno de Sá Oliveira, Centro de Pesquisas Gonçalo Mon.

Manufacturer protocol

Download the product protocol from Geraldo Gileno de Sá Oliveira, Centro de Pesquisas Gonçalo Mon for pFastBac1-sccaIL-12 below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - Hi5 IL12 canine using pFastBac1-sccaIL-12 from Geraldo Gileno de Sá Oliveira, Centro de Pesquisas Gonçalo Mon. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.