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Coomassie staining is able to detect protein bands containing about 0.2 μg or more protein. For low abundant protein, silver staining (www/silver staining) is a better choice since it is about 100-fold more sensitive than Coomassie staining.

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[Bio101] Coomassie Blue Staining
[Bio101] 考马斯亮蓝染色

生物化学 > 蛋白质 > 电泳
作者: Fanglian He
6/5/2011, 33072 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.78

[Abstract] Coomassie staining is able to detect protein bands containing about 0.2 μg or more protein. For low abundant protein, silver staining (www/silver staining) is a better choice since it is about 100-fold more sensitive than Coomassie staining.
Keywords: Coomassie staining(考马斯亮蓝染色), Coomassie blue(考马斯亮蓝), Staining solution(染色液), Destaining solution(脱色液), Protein(蛋白质)

[Abstract] 考马斯亮蓝染色能够检测出大约0.2 μg 或以上蛋白.对于低丰度蛋白银染(www/silver staining)是更好的选择,因为它比考马斯亮蓝染色敏感100倍以上.

Materials and Reagents

  1. Coomassie Brilliant Blue R250 (EM Science)
  2. Glacial acetic acid
  3. MetOH
  4. Staining solution
  5. Dye solution
  6. Destaining solution

Equipment

  1. Shaker

Procedure

  1. Incubate the gel in staining solution with shaking for 30 min or longer (can leave it overnight).
  2. Remove the dye solution (it can be reused for many times) and rinse the gel with water 1-2 times to remove the dye.
  3. Add destaining solution to the gel and incubate for 30-60 min.
  4. Transfer the gel to water (can keep it in water for several days).

Recipes

  1. 100 ml staining solution
    Coomassie Brilliant Blue R250 0.25 g
    Glacial acetic acid 10 ml
    MetOH: H2O (1: 1 v/v) 90 ml
  2. Destaining solution
    Destaining solution is the same as staining solution, but not containing the Coomassie R250 dye powder.

材料和试剂

  1. 考马斯亮蓝R250(EM Science)
  2. 冰醋酸
  3. MetOH
  4. 染色溶液
  5. 染料溶液
  6. 脱色解决方案

设备

  1. 摇床

程序

  1. 孵育凝胶在染色溶液中摇动30分钟或更长时间(可以离开过夜)
  2. 取出染料溶液(可重复使用多次),并用水冲洗凝胶1-2次,以除去染料。
  3. 添加脱色溶液到凝胶,孵育30-60分钟。
  4. 将凝胶转移到水中(可以在水中保存几天)。

食谱

  1. 100ml染色溶液 考马斯亮蓝R250 0.25 g
    冰醋酸10ml
    甲醇:H 2 O(1:1v/v)90ml
  2. 脱色解决方案
    脱色溶液与染色溶液相同,但不含有考马斯R250染料粉末
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How to cite this protocol: He, F. (2011). Coomassie Blue Staining. Bio-protocol Bio101: e78. DOI: 10.21769/BioProtoc.78; Full Text



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