title: A rapid and simple 8-quinolinol-based fluorescent stain of phosphoproteins in polyacrylamide gel after electrophoresis.
abstract: In order to obtain an easy and rapid protocol to visualize phosphoproteins in SDS-PAGE, a fluorescent detection method named 8-Quinolinol (8-Q) stain is described. 8-Q can form ternary complexes in the gel matrix contributed by the affinity of aluminum ion (Al(3+) ) to the phosphate groups on the proteins and the metal chelating property of 8-Quinolinol, exhibiting strong fluorescence in ultraviolet light. It can visualize as little as 4∼8 ng of α-casein and β-casein, 16∼32 ng of ovalbumin and κ-casein which is more sensitive than Stains-All but less sensitive than Pro-Q Diamond. The protocol of 8-Q requires only 70 min in 0.75 mm mini-size or 1.0 mm large-size gels with five changes of solutions without destaining step; Pro-Q takes at least 250 min with 11 changes of solutions. In addition, the new method was confirmed by the study of dephosphorylation and LC-MS/MS, respectively. The approach to visualize phosphoprotein utilizing 8-Q could be an alternative to simplify the analytical operations for phosphoproteomics research.
authors.affiliation: School of Pharmaceutical Sciences, Key Laboratory of Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Pharmacy & Research Institute of Drug Development, Chonnam National University, Gwangju, South Korea.
authors.forename: Sun-Young
authors.initials: SY
authors.lastname: Hwang
authors.affiliation: School of Pharmaceutical Sciences, Key Laboratory of Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors.forename: Wei-Tao
authors.initials: WT
authors.lastname: Cong
authors.affiliation: School of Pharmaceutical Sciences, Key Laboratory of Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China.
authors.forename: Li-Tai
authors.initials: LT
authors.lastname: Jin
authors.affiliation: College of Pharmacy & Research Institute of Drug Development, Chonnam National University, Gwangju, South Korea.
authors.forename: Jung-Kap
authors.initials: JK
authors.lastname: Choi
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D013058:Mass Spectrometry; D015125:Oxyquinoline / Q000737:chemistry*; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D005456:Fluorescent Dyes; D010750:Phosphoproteins; D015125:Oxyquinoline
keywords: 8-Quinolinol; Aluminum ion; Fluorescent detection; Phosphoprotein stain; SDS-PAGE