Development of a simplified, economical polyacrylamide gel staining protocol for phosphoproteins.
Pro-Q Diamond (Pro-Q DPS) is a commercially available stain that binds the phosphate moiety of phosphoproteins with high sensitivity and linearity. To conserve consumable costs we demonstrate that threefold diluted Pro-Q DPS offers the same sensitivity and linearity of signal to that obtained with undiluted Pro-Q DPS. The optimal conditions for Pro-Q DPS indicate that fixation, staining, and destaining of gels longer than 1 h, 2 h, and four 30-min incubations, respectively, are not required. The fixation and destaining solutions, but not the threefold diluted Pro-Q DPS, can be re-used without compromising the signal intensity or linear dynamic range. This modified protocol of Pro-Q DPS reduces the cost at least by fourfold, making the stain economically attractive for large-scale analysis of phosphoproteins.
Agrawal
Ganesh Kumar
GK
Biochemistry Department, University of Missouri-Columbia, 109 Life Sciences Center, Columbia, MO 65211, USA.
Thelen
Jay J
JJ
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D010750:Phosphoproteins / Q000302:isolation & purification*; D019346:Sodium Acetate / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D010750:Phosphoproteins; D019346:Sodium Acetate