Horizontal Compression on SDS Page Gel: Improving Protein Visualization with Colloidal CBB-G Staining

Colloidal Coomassie Brilliant Blue-G (CBB-G) staining has emerged as the most sensitive method for visualizing proteins on polyacrylamide gels. This method is widely used due to its compatibility with mass spectrometry (MS) analysis, sensitivity, reproducibility, and simplicity. The addition of a fixation step to the standard colloidal CBB-G staining method greatly increases protein band resolution.

The chemical structure of CBB G-250 differs from CBB R-250 by two additional methyl groups. This dye has two variants: CBB R-250, which exhibits a reddish tint, and CBB G-250, which shows a greenish tint. The anionic CBB molecules bind to proteins through hydrophobic interactions with aromatic groups and positively charged basic arginine and lysine residues.

The standard colloidal CBB-G staining method has some drawbacks, including the diffusion of proteins during the washing step, which compromises protein band resolution. To address this issue, incorporating a fixation step into the standard colloidal CBB-G staining protocol is an effective way to improve protein visualization in SDS-PAGE. This modification is fast, flexible, and retains all the advantages of the standard method.

The addition of a fixation step prevents the diffusion of proteins during the washing step, resulting in increased protein band resolution. Results show that this modification greatly increases the resolution of protein bands, making it an essential step for accurate protein visualization on SDS-PAGE gels.

In conclusion, colloidal CBB-G staining is a widely used and effective method for visualizing proteins on polyacrylamide gels. The addition of a fixation step to this method improves protein band resolution, making it an essential protocol for any proteomic study. By incorporating this modification into the standard colloidal CBB-G staining procedure, researchers can achieve more accurate results with improved protein visualization.

Horizontal compression on sds page gel