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Rapid detection of the main human plasma glycoproteins by two‐dimensional polyacrylamide gel electrophoresis lectin affinoblotting
Author(s) -
Golaz Olivier,
Gravel Patricia,
Walzer Claude,
Turler Hans,
Balant Luc,
Hochstrasser Denis F.
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601197
Subject(s) - glycoprotein , glycan , lectin , sialic acid , transferrin , chemistry , polyacrylamide gel electrophoresis , biotinylation , biochemistry , orosomucoid , gel electrophoresis , haptoglobin , membrane glycoproteins , fetuin , microbiology and biotechnology , chromatography , biology , immunology , enzyme
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 × 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 αl‐antitrypsin, αl‐antichymotrypsin, α2‐HS glycoprotein, αl‐acid glycoprotein, haptoglobin β‐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.