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. 2021 May 5;30:e00626. doi: 10.1016/j.btre.2021.e00626

Fig. 2.

Fig. 2

Expression and purification of hEKL C112S. (a) Time-profiles of cell growth and activity of hEKL C112S in flask culture. (b) SDS-PAGE analysis of flask culture samples. (c) Chromatogram of hEKL C112S purification. The inlet indicates SDS-PAGE of each fraction (raw: load fraction, UB: unbounded fraction). (d) Indirect conformation of enzymatic activity of each eluted fraction. MBP-D4K-hEKL (25 μg) was treated with 1 μl of purified hEKL, and incubated at 37 °C for 1 h. (e) SDS-PAGE and western blot of purified hEKL C112S. Enzymatic activity of hEKL C112S (f) before purification and (g) after purification. MBP-D4K-hEKL (25 μg) was treated with 1 μl of diluted culture supernatant or purified hEKL, and incubated at 37 °C for 1 h. M, Protein marker; Cont., MBP-D4K-hEKL.