S-Trap™ sample processing begins with strong, 5% SDS lysis and solubilization. This solution dissolves even poorly-soluble proteins like membrane proteins which are often left behind in the pellet. Reduction and alkylation are also performed in 5% SDS precluding any precipitation at this step. Proteins are further denatured by acidification to pH 1 to ensure complete destruction of all enzymatic activity (e.g. of proteases and phosphatases) and to maximize sensitivity to proteolysis.
Samples are then diluted with S-Trap™ binding buffer and loaded on the S-Trap™ column where proteins are captured within the submicron pores of the three-dimensional trap. Proteins captured within the trap present exceptionally high surface area, allowing them to be washed free of even pernicious contaminants (PEG, glycerol, detergents, salts, etc.). Protein capture and clean up takes only minutes.
Upon addition of your choice of protease, physical confinement of the substrate and protease within the pores of the trap forces fast digestion: the protease is either digesting your substrate or reflected off the sidewalls straight back to the protein to digest.
- S-Trap™ technology enables fast (1 hr), reproducible and universal sample processing for bottom-up proteomics. It is simple, can be performed in any lab with standard equipment and works with your choice of protease.
- S-Trap™ processing combines strong SDS-based protein solubilization with sample cleanup and rapid reactor-type protein digestion. Contaminants such as PEG, detergents, salts and glycerol are fully removed.
- S-Trap™ units are available to the multiple milligram scale. Large scale is particularly useful for those doing pre-enrichment for example in phosphopeptide or SISCAPA/immunoaffinity analysis.
- Sample processing is unaffected by protein solubility: cytosolic and membrane proteins are identically solubilized and processed.
- S-Trap™ processing is suitable for all sample types including cells, tissues, membranes and serum. With serum, 8 M urea and sample dilution is not needed.
- S-Trap™ sample processing is fast: removal of SDS and contaminants takes only few minutes, and complete digestion is achieved in as little as 30 min.
- S-Trap™ processing integrates sample digestion and peptide clean-up in the same device. It is also capable of processing sub microgram levels of protein with low losses.
- S-Traps™ also filter samples (no clogging from on-bead IP digestions, etc.)
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