Higher resolution protein band visualisation via improvement of colloidal CBB-G staining by gel fixation

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, se...

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Published inChemical and biological technologies in agriculture Vol. 9; no. 1; pp. 1 - 9
Main Authors Chong, Nikson Fatt-Ming, Hussain, Hasnain, Hamdin, Nur Ezzati, Wee, David Hong-Sheng, Nisar, Mehvish, Yan, Wei-Jie, Lau, Benjamin Yii Chung, Rahmad, Norasfaliza
Format Journal Article
LanguageEnglish
Published Cham Springer International Publishing 11.05.2022
Springer Nature B.V
SpringerOpen
Subjects
Agriculture
Biomedical and Life Sciences
Colloidal
Colloiding
Coomassie
Diffusion
Fixation
Gel electrophoresis
Gel fixation
Life Sciences
Mass spectrometry
Mass spectroscopy
Methodology
Organic Chemistry
Plant Biochemistry
Plant Physiology
Protein staining
Proteins
Proteomics
SDS-PAGE
Sodium lauryl sulfate
Soil Science & Conservation
Staining
Visualization
Washing
SDS-PAGE
Gel fixation
Colloidal
Protein staining
Proteomics
Coomassie
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Summary: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
ISSN:2196-5641
2196-5641
DOI:10.1186/s40538-022-00297-0