Not sure which gel staining method works best with MS analysis?
You can use a range of gel staining methods for Protein Identification by mass spectrometry. However, silver staining is the most sensitive staining method for low protein amounts.
The MS compatible silver stains have been optimized for compatibility and sensitivity with trypsin digestion and MS analysis*. Many staining protocols are not compatible, due to crosslinking and fixing within the gel. Many protocols do not work with mass spectrometric protein identification!
We strongly recommend that you use the two commercially available kits:
- ProteoSilver Plus, Sigma (Product # PROTSIL1 or PROTSIL2).
- Dodeca Silver Stain, BioRad (Product # 161-0481 or 161-0480).
And the Alphalyse Silver Staining protocol:
- Download the Alphalyse silver stain protocol.
The three procedures have been tested thoroughly in combination with MS protein identification. They give the best data and most sensitive protein identifications.
Alternative gel staining methods include:
- Coomassie Blue based methods. They give very good mass spectrometric data and confident protein identifications.
- Sensitive fluorescent staining methods. Such as DIGE, Deep Purple, Oriole and Sypro Ruby.
* Improved silver staining protocols for high sensitivity protein identification using matrix-assisted laser desorption/ionization-time of flight analysis. Mortz E, Krogh TN, Vorum H, Görg A. Proteomics. 2001 Nov;1(11):1359-63.
* Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Shevchenko, A, Wilm, M., Vorm, O., and Mann, M. Anal. Chem. 1996, 68, 850-858.