Development of a simplified, economical polyacrylamide gel staining protocol for phosphoproteins
Development of a simplified, economical polyacrylamide gel staining protocol for phosphoproteins
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DOI：
10.1002/pmic.200500021
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发表时间：
2005-12-01
期刊：
影响因子：
3.4
通讯作者：
Thelen, JJ
中科院分区：
文献类型：
--
作者：
Agrawal, GK;Thelen, JJ
Pro-Q Diamond （Pro-Q DPS）是一种市售染色剂，具有高灵敏度和线性，可结合磷酸蛋白的磷酸部分。为了节省耗材成本，我们证明了三倍稀释的Pro-Q DPS提供与未稀释的Pro-Q DPS相同的灵敏度和线性信号。Pro-Q DPS的最佳条件是不需要固定、染色和染色时间分别超过1小时、2小时和4次30分钟的孵育。固定和染色溶液，但不是三倍稀释的Pro-Q DPS，可以重复使用，而不会影响信号强度或线性动态范围。这种改良的Pro-Q DPS方案将成本降低了至少四倍，使该染色剂在大规模分析磷酸化蛋白方面具有经济吸引力。
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.