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. 2001 Mar 27;98(8):4776–4781. doi: 10.1073/pnas.081072398

Figure 2.

Figure 2

Expression and purification of Dslo-C280. (A) Immunoblot detection of Dslo-C280 band by anti-FLAG M2 antibody in various fractions of E. coli before (lane 1) or after (lanes 2–6) induction with IPTG. Lanes: 1, whole cells before IPTG; 2, insoluble protein pellet from lysed cells; 3, soluble protein from lysed cells; 4, whole cells; 5, soluble fraction after high sucrose treatment; and 6, soluble fraction after osmotic shock. (B) Purification via FLAG epitope on M2 anti-FLAG affinity column. SDS/PAGE of various fractions. Lanes: 1, molecular mass markers (K = kDa); 2, crude soluble extract; 3, crude inclusion body sample; 4, fraction from soluble extract after M2 column purification; and 5, fraction of renatured inclusion protein after M2 column purification. (C) Purification by SEC. SDS/PAGE of fractions from TSK G3000SW column. Lanes: 1, molecular mass markers; 2, SEC peak fraction at ≈68 kDa; and 3, SEC peak fraction at ≈34 kDa. (D) Purification via His6-tag of renatured inclusion protein. SDS/PAGE of various fractions from Ni2+-NTA column. The arrow in B, C, and D points to the Dslo-C280 band.