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Fig. 4 | BMC Research Notes

Fig. 4

From: Optimization of canine interleukin-12 production using a baculovirus insect cell expression system

Fig. 4

Analysis of purified rsccaIL-12L produced in the baculovirus-insect cell system by SDS–PAGE and Western blot. Recombinant sccaIL-12L was produced by High-five cells infected with the AcBacΔCC-sccaIL-12opt baculovirus construct at MOI 5 for 48 h. Clarified and buffer-exchanged cell culture supernatant was applied to Ni-Sepharose affinity chromatography column and the recombinant protein was eluted using an imidazole linear gradient. The chromatographic fractions were analyzed by SDS–PAGE and Western blot. Polyacrylamide gel showing: molecular weight markers (lane 1), clarified buffer-exchanged cell culture supernatant applied to the chromatographic column (10 μL, lane 2), flow through fraction (10 μL, lane 3), and sample of a chromatographic fraction containing rsccaIL-12 (10 μL, lane 4) (a). Nitrocellulose membrane showing purified sccaIL-12 incubated with normal mouse serum diluted 1:500 (lane 1) or mouse anti-his C-Term antibodies (lane 2), followed by anti-mouse immunoglobulin-alkaline phosphatase conjugate and substrate (b)

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