期刊
CHEMICAL AND BIOLOGICAL TECHNOLOGIES IN AGRICULTURE
卷 9, 期 1, 页码 -出版社
SPRINGER
DOI: 10.1186/s40538-022-00297-0
关键词
SDS-PAGE; Protein staining; Colloidal; Coomassie; Gel fixation; Proteomics
资金
- Ministry of Education Malaysia FRGS Grant [FRGS/1/2017/STG04/UNIMAS/02/1]
- Universiti Malaysia Sarawak PhD Study Fund [F07(DPP38)/1256/2015(13)]
This article describes an improved colloidal CBB-G staining method that enhances the resolution of protein bands by adding a fixation step. The modification is fast, flexible, and improves protein visualization in SDS-PAGE.
Background: Gel staining is a crucial step that allows the visualisation of proteins separated through SDS-PAGE. Colloidal Coomassie Brilliant Blue-G (CBB-G) staining is among the commonly used visualisation methods due to several factors such as compatibility with mass spectrometry (MS) analysis, sensitivity, reproducibility, and simplicity of the staining process. However, the standard colloidal CBB-G staining has a drawback: the resolution of protein bands is compromised because of diffusion of proteins during the washing step. Results: A modification to an established colloidal CBB-G staining method, which greatly increases the resolution of protein bands, is described. The addition of a fixation step, which prevents the diffusion of proteins during the washing step, is shown to increase protein band resolution. Conclusion: The fixation step is fast, flexible, and also retains all the advantages of the standard colloidal CBB-G staining methods. As there are no drawbacks, incorporating this fixation step into the standard colloidal CBB-G staining is an easy way to improve protein visualisation in SDS-PAGE.
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