FIG. 2.
SDS-PAGE, isoelectric focusing, and gel filtration analyses of CLRI. (A) Purified wild-type and recombinant CLRIs were analyzed by SDS-12% PAGE (top), and purified proteins were also analyzed by isoelectric focusing (pH 3.0 to 10.0) (bottom). Left lane, molecular weight standards; right lane, purified CLRI. (B) Pooled fractions showing RI activity from the previous step were applied to Superdex 75 calibrated with the standard proteins (Bio-Rad). Peak 1, vitamin B12 (1.359 kDa); peak 2, myoglobin (17 kDa); peak 3, ovalbumin (44 kDa); peak 4, globulin (158 kDa); peak 5, thyroglobulin (670 kDa).