Improved method for amino acid analysis of stained collagen bands from polyacrylamide gels using high-performance liquid chromatography.
A procedure is described for the determination of the amino acid composition of stained collagen bands separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis using high-performance liquid chromatography and post-labelling with o-phthalaldehyde. Six alpha-chain bands from collagens I, III and V were analysed directly after gel electrophoresis, Coomassie blue staining and destaining. It was shown that good accuracy and reproducibility can be attained using this method, which makes it possible to determine simultaneously nineteen amino acids: Asx, Thr, Ser, Glx, Pro, Cys, Gly, Ala, Val, Met, Ile, Leu, Tyr, Phe, His, Lys and Arg together with 4-Hyp and Hyl, specific amino acids found in collagen.
Hashimoto
Y
Y
Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan.
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003094:Collagen / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D006901:Hydroxylysine / Q000032:analysis; D006909:Hydroxyproline / Q000032:analysis; D008969:Molecular Sequence Data
D006901:Hydroxylysine; D003094:Collagen; D006909:Hydroxyproline