陆彩玲, 唐深, 刘楠楠, 黄玲, 李习艺, 郭松超. 海马组织蛋白不同染色方法的比较[J]. 环境与职业医学, 2013, 30(1): 26-28.
引用本文: 陆彩玲, 唐深, 刘楠楠, 黄玲, 李习艺, 郭松超. 海马组织蛋白不同染色方法的比较[J]. 环境与职业医学, 2013, 30(1): 26-28.
LU Cai-ling , TANG Shen , LIU Nan-nan , HUANG Ling , LI Xi-yi , GUO Song-chao . Comparison of Different Staining Methods for Hippocampal Proteomic Analysis[J]. Journal of Environmental and Occupational Medicine, 2013, 30(1): 26-28.
Citation: LU Cai-ling , TANG Shen , LIU Nan-nan , HUANG Ling , LI Xi-yi , GUO Song-chao . Comparison of Different Staining Methods for Hippocampal Proteomic Analysis[J]. Journal of Environmental and Occupational Medicine, 2013, 30(1): 26-28.

海马组织蛋白不同染色方法的比较

Comparison of Different Staining Methods for Hippocampal Proteomic Analysis

  • 摘要: 目的 通过实验,比较不同染色方法对海马组织蛋白质双向凝胶电泳(2-dimensional gel electrophoresis,2-DE)分离效果的影响,为类似的海马组织蛋白分离研究提供参考。

    方法 分离、制备大鼠海马组织总蛋白样品并进行蛋白定量,不同含量海马蛋白质经2-DE分离后分别采用改良考马斯亮蓝染色、硝酸银染色法,中浓度上样量的的凝胶经考马斯亮蓝染色后也用考马斯亮蓝-硝酸银进行复染,比较不同染色方法对2-DE分离效果的影响。

    结果 不同染色方法随上样量增加,所见蛋白斑点增加,干扰物质也增加。在本研究条件下,700 μg上样量采用硝酸银染色效果最佳,考马斯亮蓝则显示1 800 μg上样量效果更优。1 200 μg考马斯亮蓝-硝酸银复染的效果较单独考染效果好。

    结论 硝酸银染色较改良考马斯亮蓝更敏感,凝胶所见蛋白质斑点更多,考马斯亮蓝-硝酸银复染效果较单独改良考马斯亮蓝好。

     

    Abstract: Objective To compare different staining methods for hippocampal proteomic analysis using 2-dimensional gel electrophoresis (2-DE) and to provide basis for similar researches.

    Methods Rat hippocampus were isolated and prepared for total protein samples. Supernatant was determined by Bradford method and separated by 2-DE. Modified Coomassie brilliant blue staining and silver staining were employed for the detection of total protein loaded in samples of different hippocampus protein levels. Specially, Coomassie brilliant blue-silver staining was applied again for the medium loading sample.

    Results With the loading protein increasing, the protein spots and impurities were increasing. In the present study, silver staining was ideal for the loading amount of 700 μg protein and modified Coomassie brilliant blue staining for 1 800 μg. Coomassie brilliant blue-silver dual staining was superior to Coomassie brilliant blue staining alone for the 1 200 μg loading sample.

    Conclusion More spots can be observed after silver staining which is more sensitive to 2-DE hippocampus separation than modified Coomassie brilliant blue staining. The sensitivity of Coomassie brilliant blue-silver dual staining is better than single modified Coomassie brilliant blue staining.

     

/

返回文章
返回