Enzyme purity, western blot and molecular mass analysis of SmIDH.

Abstract

<p>(A) SDS-PAGE analysis of the expression and purification of SmIDH. Analysis was performed on 12% polyacrylamide gel. M, protein markers; lane 1, crude extracts of cells harboring pWT grown in MD medium without glutamate. lane 2, purified protein. (B) Western blot analysis using anti-6×His antibody as probe. (C) Molecular mass determination by gel filtration chromatography. The flow rate was 0.5 ml/min and the proteins in the fractions were monitored at 280 nm. Inside is the standard curve for molecular mass. SmIDH was represented as a dark circle (•). Standard proteins were represented by open circles (○): 1, Ovalbumin (44 kDa); 2, Conalbumin (75 kDa); 3, Aldolase (158 kDa); 4, Ferritin (440 kDa); 5, Thyroglobulin (669 kDa). <i>V</i>_e of SmIDH is 14.4 ml.</p