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. 2011 Sep;193(17):4523–4530. doi: 10.1128/JB.00231-11

Fig. 4.

Fig. 4.

(A) Spin-down assay of Ccrp58 after purification from E. coli. Coomassie-stained SDS-PAGE of spin-down assays of 20-μl fractions. S, supernatants; P, pellets resuspended in 20 μl SDS loading buffer. Different pH levels are indicated. (B) Western blot of 20 μg of whole-cell extracts of H. pyloriwild-type cells (KE) and of the mutant, in which Ccrp58 is fused to the Strep tag (KE-58Strep) as indicated. M, marker; marker protein sizes are indicated. The antiserum recognizes the tagged version of Ccrp58 in strain KE-58Strep, as well as a nonspecific band, which can also be seen with the parent strain KE. (C to F) Analytical gel filtration of Ccrp proteins. Elution profiles of analytical gel filtration of Ccrp58 (C) and Ccrp1142 (D) using a Superose6 10/300 column (left) and a Biosep-SEC-S4000 column (right). In the right panels, the red curves display gel filtration immediately after purification, the blue curves after 2 weeks, and the green curves after 4 weeks after purification. Elution profiles of Ccrp59 (E) and Ccrp1143 (F) using a Superose6 10/300 column. The triangles above each elution profile indicate the elution times of the standard proteins (Bio-Rad) as follows: thyroglobulin (670 kDa, 19 S), bovine gamma-globulin (158 kDa, 7.4 S), chicken ovalbumin (44 kDa, 3.5 S), equine myoglobin (17 kDa, 2.0 S), vitamin B12(1.35 kDa). mAU, milliabsorbance units.