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Fig. 1 | BMC Biotechnology

Fig. 1

From: Preparation of an anti-NEK2 monoclonal antibody and its application in liver cancer

Fig. 1

Cloning and Expression of NEK2 in prokaryotic system. a The construction of the pET30a-NEK2 vector. M: DNA marker, 1: PCR amplification product of the NEK2 gene, 2: Colony PCR detection. b BamHI and Sa1I enzymatic digestion of the recombinant plasmid. c Expression of the recombinant human NEK2 protein. M: marker, 1: Precipitate of the pET30a group with induction, 2: Precipitate of the pET30a group without induction, 3: Precipitate of the pET30a-NEK2 group with induction, 4: Precipitate of the pET30a-NEK2 group without induction, 5: Supernatant of the pET30a group with induction, 6: Supernatant of the pET30a group without induction, 7: Supernatant of the pET30a-NEK2 group with induction, 8: Supernatant of the pET30a-NEK2 group without induction. d Expression of recombinant human NEK2 protein at different temperatures. M: marker, 1: pET30a was induced, 2: pET30a was not induced, 3: pET30a-NEK2 was not induced, 4–7: pET30a -NEK2 was induced at 18 °C, 28 °C, 37 °C, and 42 °C. e Expression of recombinant human NEK2 protein at different IPTG concentrations. M-3: The same as the (d), 4–8: pET30a-NEK2 was induced with 0.2 mmol/L, 0.4 mmol/L, 0.6 mmol/L, 0.8 mmol/L, and 1.0 mmol/L IPTG. f Expression of recombinant human NEK2 protein at different times. M-3: The same as the (d), 4–12: pET30a -NEK2 was induced at 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, and 36 h. g SDS-PAGE analysis of recombinant human NEK2 protein purified. h Western blot analysis of purified NEK2 protein using an anti-His monoclonal antibody. i Mass spectrometry identification of the recombinant human NEK2 protein with Mascot alignment. For improved clarity and conciseness, cropped areas of the blot are shown. The full length (uncut) blot image is shown in Additional file 2: Fig. S1–S8, respectively

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