STAINING OF 1D/2D SDS PAGE FOR PROTEIN ID
You can use a broad range of gel staining procedures when preparing samples for the Pick ‘n Post Protein Identification.

In general, different Coomassie Blue staining methods and commercial Coomassie staining kits gives very good mass spectrometric data and confident protein identifications.

If you need more sensitive staining methods due to lower protein amounts, most fluorescence staining methods can also be used, for example DIGE stains, Deep Purple and Sypro Ruby.

For low protein amounts we recommend to use silver staining methods that have been optimized for compatibility and sensitivity with trypsin digestion and MS analysis. Many silver staining protocols are not compatible and do not work with mass spectrometric protein identification!

You can either use one of the two commercially available kits mentioned below or download our protocol below. The three procedures have been tested thoroughly in our laboratory in combination with MS protein identification.
Commercially Available Silver Staining kits:

• ProteoSilver Plus, Sigma (Product # PROTSIL1 or PROTSIL2)
• Dodeca Silver Stain, BioRad (Product # 161-0481 or 161-0480)

“Home-made” Silver Staining

• Pick 'n Post Silver Staining Protocol

1D/2D PVDF MEMBRANES FOR N-TERMINAL SEQUENCING
For N-terminal Edman sequencing SDS PAGE separated proteins can be electroblotted onto PVDF membranes, the membranes stained and protein bands/spots cut out for N-terminal Edman sequencing. Compatible PVDF staining methods include Ponceau S, Coomassie Blue, Amido Black, and fluorescence Sypro Ruby staining. Silver staining and Western blots cannot be used in combination with Edman sequencing! You can download our protocols for electroblotting and PVDF staining below.

• Pick ‘n Post PVDF Blotting and Staining Protocols