Name: Common Issues and Solutions in Coomassie Blue Staining
Creator: AAT Bioquest
License: https://www.aatbio.com/tou.html

Common Issues and Solutions in Coomassie Blue Staining

Problem	Possible Reason	Troubleshooting
Reproducibility is difficult	Incubation timings and agitation might not have been uniform.	Maintain consistent timing and agitation each time by documenting exact conditions.
Bands are too faint/ Poor staining	Insufficient amount of protein. Gel is not fixed properly. Under-staining. Over-destaining.	Optimize the protein concentration needed to run on the gel for proper visualization. Use protein standards if needed. Each step holds significance, ensure proper completion of each step during entire staining process.
Unclear gels	Insufficient protein separation during electrophoresis.	Ensure completion of protein separation on gels by monitoring the movement of dye front.
Uneven staining	Uneven distribution of solutions (fixing/staining/de-staining/washing) on the gel.	Gentle but thorough agitation of the gel during incubation making sure that the gel is completely covered with solution.
Blotch marks	Uneven fixing. Contaminants present in the sample/reagents. Mishandling/Over-handling of gels.	Fixing step should be kept uniform with proper agitation. Use high-quality reagents and clean glassware for preparing and handling solutions. Avoid touching the gel as far as possible. If handling, always wear gloves.
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References

This online resource may be cited as follows

MLA	"Quest Database™ Common Issues and Solutions in Coomassie Blue Staining." AAT Bioquest, Inc., 6 Dec. 2025, https://www.aatbio.com/data-sets/common-issues-and-solutions-in-coomassie-blue-staining.
APA	AAT Bioquest, Inc. (2025, December 6). Quest Database™ Common Issues and Solutions in Coomassie Blue Staining. AAT Bioquest. https://www.aatbio.com/data-sets/common-issues-and-solutions-in-coomassie-blue-staining.
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