Tritium/PPO gel fluorographic efficiency is reduced by Coomassie Blue staining
Tritium/PPO gel fluorographic efficiency is reduced by Coomassie Blue staining
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考马斯蓝染色会降低氚/PPO 凝胶荧光效率
DOI：
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发表时间：
1982
期刊：
影响因子：
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通讯作者：
M. Dahmus
中科院分区：
文献类型：
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作者：
R. C. Higgins;M. Dahmus
当用聚丙烯酰胺凝胶电泳法分析[~3H]标记的蛋白质时，如果凝胶被考马斯亮蓝R-250染色，用PPO(2，5-二苯基恶唑)浸渍凝胶后的荧光检测灵敏度显著降低。由于荧光照相效率的降低取决于着色剂的浓度，而着色剂的浓度又取决于蛋白质的浓度，因此根据样品组成的不同，荧光照相的定量测量可能会产生很大的误差。当允许甲醇在DMSO和DMSO/PPO溶液中积聚时，由于凝胶对PPO的渗透性降低，导致荧光照相效率的另一种降低。然而，这种荧光照相灵敏度的损失应该与样品无关。
When [3H]‐labeled proteins are analyzed by polyacrylamide gel electrophoresis, the sensitivity of fluorograhic detection following gel impregnation with PPO (2,5‐diphenyloxazole) in DMSO (dimethylsulfoxide) is significantly reduced if the gel has been stained with Coomassie Brilliant Blue R‐250. Since the reduction in fluorographic efficiency depends on the concentration of stain, which in turn depends upon protein concentration, substantial errors may enter into quantitative measurements of fluorograms depending on sample composition. Another reduction in fluorographic efficiency results from reduced penetrability of the gels to PPO when a build‐up of methanol in DMSO and DMSO/PPO solutions is allowed. This loss of fluorographic sensitivity should be sample independent, however.