Simple quantification of Cu,Zn-superoxide dismutase activity after separation by non-denaturing isoelectric focusing.
The Cu,Zn-superoxide dismutase (SOD) activity in bovine retina cytosol was separated from retinal pigment using short-length non-denaturing isoelectric focusing (IEF) (15-mm long x 1.3-mm i.d. column) and detected using non-denaturing two-dimensional electrophoresis (2-DE). After the SOD and pigment in the retina cytosol are separated, SOD activity can be quantified by water-soluble tetrazolium salt. We also found that SOD separated by this IEF retained its native function.
Shimazaki
Youji
Y
Department of Material Science, Faculty of Science and Venture Business Laboratory, Ehime University, Matsuyama City, 790-8577 Japan. yoji@dpc.ehime-u.ac.jp
Ohnishi
Hiromi
H
Matsuura
Satoshi
S
Manabe
Takashi
T
D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation / Q000502:physiology; D007525:Isoelectric Focusing / Q000379:methods*; D010449:Peptide Mapping; D015203:Reproducibility of Results; D012160:Retina / Q000737:chemistry* / Q000201:enzymology*; D012168:Retinal Pigments / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000032:analysis / Q000737:chemistry*; D013778:Tetrazolium Salts / Q000737:chemistry
D012168:Retinal Pigments; D013778:Tetrazolium Salts; D013482:Superoxide Dismutase