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.
