Author:
Chong Nikson Fatt-Ming,Hussain Hasnain,Hamdin Nur Ezzati,Wee David Hong-Sheng,Nisar Mehvish,Yan Wei-Jie,Lau Benjamin Yii Chung,Rahmad Norasfaliza
Abstract
Abstract
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.
Graphical Abstract
Funder
Universiti Malaysia Sarawak PhD Study Fund
Ministry of Education Malaysia FRGS Grant
Universiti Malaysia Sarawak
Publisher
Springer Science and Business Media LLC
Subject
Agronomy and Crop Science,Biochemistry,Food Science,Biotechnology
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