Detection of glycoproteins in polyacrylamide gels and on electroblots using Pro-Q Emerald 488 dye, a fluorescent periodate Schiff-base stain.
Pro-Q Emerald 488 glycoprotein stain reacts with periodic acid-oxidized carbohydrate groups, generating a bright green-fluorescent signal on glycoproteins. The stain permits detection of less than 5-18 ng of glycoprotein per band, depending upon the nature and the degree of protein glycosylation, making it roughly 8-16-fold more sensitive than the standard colorimetric periodic acid-Schiff base method using acidic fuchsin dye (pararosaniline). The green-fluorescent signal from Pro-Q Emerald 488 stain may optimally be visualized using charge-coupled device/xenon arc lamp-based imaging systems or 470-488 nm laser-based gel scanners. Though glycoprotein detection may be performed on transfer membranes, direct detection in gels avoids electroblotting and the specificity of staining is better in gels. After detecting glycoproteins with Pro-Q Emerald 488 dye, total protein profiles may subsequently be detected using SYPRO Ruby protein gel stain. Using computer-assisted registration techniques, images may then be merged to generate differential display maps.
Hart
Courtenay
C
Molecular Probes, Eugene, OR 97402, USA.
Schulenberg
Birte
B
Steinberg
Thomas H
TH
Leung
Wai-Yee
WY
Patton
Wayne F
WF
D000180:Acrylic Resins / Q000737:chemistry*; D001386:Azides / Q000737:chemistry; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005456:Fluorescent Dyes / Q000737:chemistry*; D006023:Glycoproteins / Q000032:analysis*; D012545:Schiff Bases; D013050:Spectrometry, Fluorescence; D000099166:Polyacrylamides
D000180:Acrylic Resins; D001386:Azides; D005456:Fluorescent Dyes; D006023:Glycoproteins; D012545:Schiff Bases; D000099166:Polyacrylamides