Fluorescent staining of glycoproteins on polyvinylidene difluoride membrane with 8-aminonaphthalene-1,3,6-trisulfonate.
Here, we describe a simple and sensitive method that allows fluorescent detection of glycoproteins on polyvinylidene difluoride (PVDF) membrane. We used periodic acid oxidation of carbohydrate chains of glycoproteins and fluorescent labeling with 8-aminonaphthalene-1,3,6-trisulfonate (AN-TS) by reductive amination. We developed an additional method to enhance the ability of PVDF to absorb glycoproteins by using non-glycoprotein lectin, such as wheat germ agglutinin (WGA), as a link between the PVDF membrane and glycoproteins, resulting in considerably increased detection sensitivity to glycoproteins.
Che
F Y
FY
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai, 200031, PRC.
Song
J F
JF
Yu
L R
LR
Wang
K Y
KY
Xia
Q C
QC
D000042:Absorption; D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000032:analysis*; D005470:Fluorometry / Q000191:economics / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D015151:Immunoblotting; D037102:Lectins / Q000737:chemistry; D008567:Membranes, Artificial; D009281:Naphthalenes / Q000032:analysis*; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000191:economics / Q000379:methods*; D014909:Wheat Germ Agglutinins / Q000737:chemistry; D005465:Fluorocarbon Polymers
D005456:Fluorescent Dyes; D006023:Glycoproteins; D037102:Lectins; D008567:Membranes, Artificial; D009281:Naphthalenes; D010504:Periodic Acid; D011145:Polyvinyls; D014909:Wheat Germ Agglutinins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid