Fig. 3

Refolding of MucB protein on a gel filtration column. The denatured MucB protein was injected on a Superdex 200 HR 10/30 column equilibrated with the NAT refolding buffer and connected to the FPLC system. a elution profile from the column monitored by absorbance at 280 nm. b analysis of fractions eluted from the column on a 12% SDS-polyacrylamide gel stained with Coomassie Brilliant Blue R 250. The migration of molecular weight standards in the SDS-polyacrylamide gel is shown on the left. The mobility of molecular weight markers of the indicated molecular weights in the column, as determined in a separate run, is shown on the top of the gel. V₀ is void volume. The molecular weight markers are as follows: β-amylase, 200 kDa; alcohol dehydrogenase, 150 kDa; bovine serum albumin, 66 kDa; carbonic anhydrase, 29 kDa; cytochrome C, 12.4 kDa. The fractions corresponding to the column void volume and the elution volume of the collected MucB peak (eluting between 12.4 and 13.6 mls) are labeled with arrows at the bottom of the gel