How it works
Protein sample preparation is a vital first step for mass spectrometry and gel-based protein analysis.
At Impact Proteomics, we provide protein sample preparation tools to ensure your workflow begins with the highest quality sample so you can get the highest quality data.
Whether you are working with whole proteins, whole proteomes, or peptides our protein sample preparation tools can enhance your workflows using our patent pending reversible tagging system. We have taken advantage of natural protein chemistry to create a way to capture, clean up, and release any protein-based sample.
Our ProMTag reversible protein tagging agent reacts with primary amines on the surface of every protein and/or peptide to covalently link the tag to the surface. The ProMTag agent then reacts with the surface of the ProMTag Capture Resin to covalently attach the proteins to the surface of the resin, allowing for extensive washing and buffer exchange. The covalent bond between the protein and the ProMTag is reversed by adding the mass spec compatible elution buffer. This leaves the amines on the protein in their original, unmodified state ensuring no changes to the mass or isoelectric point of the protein that could interfere with analysis. If the proteins are being digested into peptides mP-trypsin, which is stabilized to prevent autodigestion and labeled with an irreversible tag to prevent contamination of the final sample, can be added.
All of our protein extraction and cleanup kits contain the same four basic steps:
- 1) Label proteins with ProMTag, our reversible protein tag
- 2) Covalently bind ProMTag labeled proteins to ProMTag capture resin
- 3) Wash away contaminants and/or exchange buffers
- 4) Release the clean, unmodified protein sample from the beads
For the generation of peptide samples for mass spectrometry, additional steps include:
- 5) Reduction and alkylation
- 6) Digestion with stabilized and tagged trypsin
- 7) Removal of trypsin
- 8) Elution of clean, digested peptides
All reagents and supplies for all steps are included in our kits for convenience and reproducibility