Higher resolution protein band visualisation via improvement of colloidal CBB-G staining by gel fxation
Background: Gel staining is a crucial step that allows the visualisation of proteins separated through SDS-PAGE. Col‑ loidal 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) analysi...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BioMed Central Ltd
2022
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/38633/1/Higher%20resolution%20-%20Copy.pdf http://ir.unimas.my/id/eprint/38633/ https://chembioagro.springeropen.com/articles/10.1186/s40538-022-00297-0 https://doi.org/10.1186/s40538-022-00297-0 |
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| Summary: | Background: Gel staining is a crucial step that allows the visualisation of proteins separated through SDS-PAGE. Col‑
loidal 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 difusion of proteins during the washing step.
Results: A modifcation to an established colloidal CBB-G staining method, which greatly increases the resolution of
protein bands, is described. The addition of a fxation step, which prevents the difusion of proteins during the wash‑
ing step, is shown to increase protein band resolution.
Conclusion: The fxation step is fast, fexible, and also retains all the advantages of the standard colloidal CBB-G stain‑ing methods. As there are no drawbacks, incorporating this fxation step into the standard colloidal CBB-G staining is an easy way to improve protein visualisation in SDS-PAGE. |
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