Research Paper Volume 1, Issue 6 pp 557—572

Figure 1. Muscleeeeee creatine kinase (CKm) purified from young (3-6 months), middle aged (12-14 months) and aged (20-24 months) mouse quadriceps. (A) Peak Blue Sepharose fractions of purified CKm (1 μg) from young (lane 1), middle aged (lane 2) and aged (lane 3) mouse muscle were resolved on a denaturing SDS gel and stained with Coomassie Blue. These fractions are ~85% purified CKm and were used for enzyme kinetic analyses. CKm within side fractions from the Blue Sepharose pH gradient elution were pooled and purified to a single band using hydroxyapatite chromatography. Lanes 4-6 represent samples from young, middle-aged and aged mice, respectively. (B) Western blot analysis of CKm levels in fractions eluted from a Blue Sepharose column, using an anti-creatine kinase type M antibody. (C) Densitometric analysis demonstrates the increase in carbonylated CKm in quadriceps of young, middle aged and aged mice. The peak level of carbonylation occurs in muscle of middle aged mice. (D) immunoblot analysis of carbonylated CKm in Blue Sepharose fractions [3-6], The carbonylated CKm was identified by anti-DNP antibody.