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Published on: December 30, 2013
Coomassie staining as loading control in Western blot analysis
Charlotte Welinder1, Lars Ekblad
1Department of Oncology, Clinical Sciences, Lund University, Sweden.
Insights
A new, faster protein staining method simplifies Western blotting analysis. This technique is compatible with standard blocking and can be used after immunodetection for improved results and further identification.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Western blotting commonly uses housekeeping proteins to normalize electrophoresis loading.
- Existing protein staining methods can be time-consuming and may interfere with subsequent analyses.
Purpose of the Study:
- To develop a faster and simpler protein staining method for Western blotting.
- To ensure compatibility with standard blocking conditions and immunodetection protocols.
- To offer a method suitable for post-immunodetection analysis with enhanced linearity.
Main Methods:
- A novel protein staining protocol was developed and tested.
- Compatibility with standard blocking conditions was evaluated.
- The method's performance after immunodetection was assessed for linearity.
Main Results:
- The new staining method is significantly faster and simpler than conventional techniques.
- It is fully compatible with ordinary blocking conditions used in Western blotting.
- Post-immunodetection staining demonstrated superior linearity compared to existing methods.
Conclusions:
- This improved protein staining technique offers a more efficient workflow for Western blotting.
- Its compatibility and superior linearity make it a valuable tool for protein analysis.
- The method allows for further identification of protein bands using peptide mass fingerprinting.
Abstract:
In Western blotting, immunodetection of housekeeping proteins is routinely performed to detect differences in electrophoresis loading. The present work describes a much faster and simpler protein staining method, which is compatible with ordinary blocking conditions. In addition, the method can be used after immunodetection with superior linearity compared to ordinary staining methods. After immunoblotting and staining, protein bands can be further identified using peptide mass fingerprinting.
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