Detection of sialylated phosphorylated kappa-casein glycomacropeptide electrophoresed on polyacrylamide gels and cellulose acetate strips by the thiobarbituric acid and malachite green dye reactions.
A 64 amino acid residue sialylated phosphorylated glycomacropeptide (GMP) from bovine sweet whey can be detected as a Coomassie blue-staining peptide by electrophoresis on sodium dodecyl sulfate (SDS)-polyacrylamide gels. There is, however, limited information available concerning detection of GMP as a sialylated phosphorylated compound. Samples of GMP were electrophoresed on SDS-polyacrylamide gels or cellulose acetate strips (CAS). Immediately following electrophoresis, fractions obtained by cutting gels or strips were subjected to sialic acid determination by the thiobarbituric acid reaction and phosphorus determination by the malachite green dye reaction. Both determinations were found to be sensitive enough to detect approximately 20 and 40 microg of GMP in CAS and SDS gels, respectively. Further studies demonstrated that sialylated phosphorylated GMP can be detected on either SDS gels or CAS loaded with whey products or whey-added margarine residues.
Nakano
Takuo
T
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5. takuo.nakano@afhe.ualberta.ca
Ikawa
Noriaki
N
Ozimek
Lech
L
D000818:Animals; D002364:Caseins / Q000032:analysis* / Q000737:chemistry*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D008892:Milk / Q000737:chemistry; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D010766:Phosphorylation
D002364:Caseins; D006020:Glycopeptides; D010758:Phosphorus; D019158:N-Acetylneuraminic Acid