How can I confirm the PTMScan® LysC Protease #84748 digested my proteins efficiently?

To confirm digestion, we recommend taking a ~1% aliquot of the protein sample pre- and post-digestion and running them on an SDS-PAGE gel with Coomassie blue staining. After digestion, you should see minimal Coomassie staining, possibly with some "fuzzy" bands visible below 11 kDa in the post-digestion sample.

Alternatively, you can continue through peptide purification and take 0.1-0.5% of the resulting eluted peptides from the Sep-Pak, dilute with 0.1% trifluoroacetic acid by at least a ten-fold volume, and analyze using liquid chromatography-mass spectrometry (LC–MS). The LC-MS file can then be analyzed to assess the number of miscleaved peptides.

Last updated: September 12, 2024