ProteoExtract - Protein Digestion

Overview:

The ProteoExtract^® All-in-One Trypsin Digestion Kit contains a set of optimized reagents that allows tryptic digests of various protein sample types: spots or bands excised from polyacrylamide gels, protein solutions, or cell and tissue extracts. Each protocol is optimized using affinity-purified trypsin, which guarantees efficient digestion of protein samples regardless of their origin. High digestion efficiency results in a high yield of tryptic peptides—even with hard-to-digest proteins—leading to improved LC-MS peak patterns and sequence coverage after MS analysis. This allows a comprehensive analysis of post-translational modifications such as phosphorylation site identification.

Features:
- Compatible with a variety of protein sample types
- Fast procedure, complete digestion in 3 hours
- 20–30% increase in sequence coverage

Application Example :
The ProteoExtract Digestion Kit generates greater MS sequence coverage
compared to a competitor product.

As given in Table 1 certain amounts of proteins were separated on a 12% Tris-glycine SDS-PAGE gel. The gel was stained with colloidal Coomassie™, the bands were excised, destained and tryptically digested with the ProteoExtract^® Trypsin Digestion Kit (2 hrs) or with a competitor product (4 hrs) according to the manufacturers’ protocols. The tryptic peptides were then analyzed by LC-MS/MS with subsequent database search using the Mascot algorithm and the SWISS-PROT/TrEMBL database

Improved digestion of hard-to-digest proteins Ovalbumin and ovalbumin digests were analyzed as follows: lane 1: ovalbumin, chicken egg, 43 kDa; lane 2: digest incubated 3 h using ProteoExtract All-in-One Trypsin Digestion Kit; lane 3: digest incubated 4 h using the same kit; lane 4: digest incubated 3 h using reagent from Supplier I; lane 5: digest incubated 3 h using kit from Supplier II.

Enhanced detection allows comprehensive analysis Ovalbumin was digested using ProteoExtract All-in-One Trypsin Digestion Kit (panel A) or according to Supplier III kit protocol (panel B). Base peak chromatograms of nano-LC/MS analyses are shown.