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Fig. 3 | BMC Biotechnology

Fig. 3

From: Design, purification and assessment of GRP78 binding peptide-linked Subunit A of Subtilase cytotoxic for targeting cancer cells

Fig. 3

Purification and refolding of GBP-SubA. a. After three washes with PBS, the irrelevant proteins of insoluble lysate fraction reduced. Lane 1 and 2: supernatant and pellet after the 1st wash; lane 3 and 4: supernatant and pellet after the 2nd wash; lane 5 and 6: the supernatant and pellet after the 3rd wash; Lane M: marker. b. GBP-SubA proteins were eluted by a gradient concentration of imidazole from nickel ion-chelated beads. Lane 1–3: the protein eluted by 50 mM imidazole; lane 4–6: the protein eluted by 100 mM imidazole; Lane 7–9: the protein eluted by 200 mM imidazole; Lane 10–12: the protein eluted by 300 mM imidazole; Lane 13–15: the protein eluted by 500 mM imidazole; Lane M: marker. c. Refolded-GBP-SubA protein was analyzed by SDS-PAGE and coomassie brilliant blue R-250 stain. Lane 1: the GBP-SubA protein after refolding; Lane M: marker. d. Refolded-GBP-SubA protein was analyzed by SDS-PAGE and silver stain. Lane 1: the GBP-SubA protein after refolding; Lane M: marker

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