Rapid detection of the main human plasma glycoproteins by two-dimensional polyacrylamide gel electrophoresis lectin affinoblotting.
Glycoprotein modifications in the glycan moiety can occur in diseases such as cancers, inflammatory processes and alcoholism. We combined high-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) with lectin affinoblotting in order to establish the normal human plasma glycoprotein map. Human plasma proteins were separated by mini 2-D PAGE (7 x 9 cm), transferred onto polyvinylidene difluoride membranes and incubated with biotinylated lectins. We focused our study on lectins binding sialic acid and galactose residues. Known plasma glycoproteins such as alpha 1-antitrypsin, alpha 1-antichymotrypsin, alpha 2-HS glycoprotein, alpha 1-acid glycoprotein, haptoglobin beta-chain and transferrin were easily detected in ng amounts. This protocol was adequate to establish a normal plasma glycoprotein map and will allow the study of glycoproteins in diseases.
D015180:Electrophoresis, Gel, Two-Dimensional; D005690:Galactose / Q000737:chemistry; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D037102:Lectins; D019158:N-Acetylneuraminic Acid; D011485:Protein Binding; D012794:Sialic Acids / Q000737:chemistry; D013997:Time Factors
D006023:Glycoproteins; D037102:Lectins; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid; D005690:Galactose