title: Modern bioanalysis of proteins by electrophoretic techniques.
abstract: In 1957, protein rich in cysteine able to bind cadmium was isolated from horse kidney and named as metallothionein according to its structural properties. Further, this protein and metallothionein-like proteins have been found in tissues of other animal species, yeasts, fungi and plants. MT is as a potential cancer marker in the focus of interest, and its properties, functions, and behavior under various conditions are intensively studied. Our protocol describes separation of two major mammalian isoforms of MT (MT-1 and MT-2) using capillary electrophoresis (CE) coupled with UV detector. This protocol enables separation of MT isoforms and studying of their basic behavior as well as their quantification with detection limit in units of ng per μL. Sodium borate buffer (20 mM, pH 9.5) was optimized as a background electrolyte, and the separation was carried out in fused silica capillary with internal diameter of 75 μm and electric field intensity of 350 V/cm. Optimal detection wavelength was 254 nm.
authors.affiliation: Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University, Brno, Czech Republic.
authors.forename: Sona
authors.initials: S
authors.lastname: Krizkova
authors.forename: Marketa
authors.initials: M
authors.lastname: Ryvolova
authors.forename: Michal
authors.initials: M
authors.lastname: Masarik
authors.forename: Ondrej
authors.initials: O
authors.lastname: Zitka
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
authors.forename: Jaromir
authors.initials: J
authors.lastname: Hubalek
authors.forename: Tomas
authors.initials: T
authors.lastname: Eckschlager
authors.forename: Rene
authors.initials: R
authors.lastname: Kizek
mesh_terms: D000595:Amino Acid Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
chemical_list: D011506:Proteins