| 电泳凝胶与转膜的原位切割方法 |
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| 引用本文： | 程熠,洪敏,高琳,李奇. 电泳凝胶与转膜的原位切割方法[J]. 吉林大学学报(医学版), 2003, 29(3): 264-266 |
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| 作者姓名： | 程熠 洪敏 高琳 李奇 |
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| 作者单位： | 1. 吉林大学基础医学院生物化学与分子生物学教研室,吉林,长春,130021
2. 吉林修正药业 |
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| 基金项目： | 吉林省计划委员会资助项目;9905G168; |
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| 摘 要： | 目的 :建立简易的快速分离高活性纯度酶以及用于氨基酸测序的转膜方法。方法 :以考马斯亮蓝 R- 2 5 0的磺基水杨酸水基染色液 ,确定聚丙烯酰胺凝胶与转印到 PVDF膜的蛋白电泳带的位置 ,以等位切割方法获得未受染色液污染的高纯度的活性酶组分以及转印的 PVDF膜。结果 :以适当浓度的考马斯亮蓝 R- 2 5 0的磺基水杨酸水基染色液染色 ,能保证聚丙烯酰胺凝胶和 PVDF膜在染色前后保持等长。结论 :等位切割方法能避免目前常见的如 SDS、染色液组分对酶活性以及测序结果造成的干扰
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| 关 键 词： | 电泳,聚丙烯酰胺凝胶/方法 转印 原位切割 |
| 文章编号： | 1671-587X(2003)03-0264-03 |
| 修稿时间： | 2002-04-28 |
In Situ Cut in Electrophoretic Gel and Transblotted PVDF Membrane |
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| Abstract: | Objective: To establish simple method in fast separation for active enzyme of high purity and transblotting for amino acid sequencing. Methods: Water based staining solution was prepared by Coomassic Brilliant Blue R 250 and sulfosalicylic acid. Protein bands and transbloting band were determined. Active enzyme of high purity from unstained electrophoretic gel and unstained transblotted PVDF membrane were obtained by in situ cut. Results:The gel and PVDF membrane stained by staining solution of suitable concentration can keep same length with unstained counterpart. Conclusion:Interference resulted from reagents was avoided by in situ cut. |
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| Keywords: | Electrophoresis Polyacrylamide/methods Transbloting In situ cut |
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