Development of a simplified, economical polyacrylamide gel staining protocol for phosphoproteins |
| |
Authors: | Agrawal Ganesh Kumar Thelen Jay J |
| |
Institution: | Biochemistry Department, University of Missouri-Columbia, 109 Life Sciences Center, Columbia, MO 65211, USA. |
| |
Abstract: | 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. |
| |
Keywords: | Fluorescent protein stain Phosphoproteins Phosphopeptides Phosphoproteome |
本文献已被 PubMed 等数据库收录! |
|