Coomassie Protein Gel Imaging

Coomassie blue protein stain is a strong 700 nm fluorophore. Its fluorescence is induced upon protein binding. A common gel stain, Coomassie blue protein stain can be used for sensitive, quantitative protein detection in gels, with a linear range from ~10 ng to 20 µg.1 It has been shown to be more sensitive than white-light imaging of Coomassie staining.1, 2

Coomassie-stained SDS-PAGE gels can be imaged on any of the Odyssey® Imagers with comparable sensitivity to Sypro Ruby, but at a lower cost.1

figure 1
Figure 1. Unstained protein standards were loaded across a 10% SDS-PAGE gel, separated by electrophoresis and stained with Coomassie Blue. The gel was imaged with the Odyssey Classic Infrared Imaging System in the 700nm channel.
figure 1
Figure 2. Mouse liver membrane and soluble proteins were separated on 2D gels, then stained with Sypro Ruby followed by Coomassie. Image analysis indicates differences in average SNR (panel G) and total spot number (panel H); * and ** denote significant differences compared to Sypro Ruby (P<0.05 and P<0.001; n=3). Reprinted from Harris, LR et al.

See some published examples of protein gel imaging

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References

  1. Luo, S. et al. (2006). Analytical Biochemistry, 350(2), pp 233-238. DOI: 10.1016/j.ab.2005.10.048
  2. Harris, L.R. et al. (2007). Journal of Proteome Research, 6(4), pp 1418-1425. DOI: 10.1021/pr0700246.

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