How it works
Sample preparation is a vital first step for mass spectrometry and gel-based protein analysis.
At Impact Proteomics, we provide sample preparation tools to ensure your workflow begins with the highest quality sample so you can get the highest quality data.
The key to our incredible sample preparation tools is our patent pending reversible click chemistry tag, called ProMTag. One half of the tag reacts reversibly with primary amines on the surface of proteins and the other half covalently reacts with ProMTag capture resin. This allows us to quickly tag proteins, immobilize them covalently on resin, wash away contaminants, and then release the proteins in their original, unmodified state.
We have taken advantage of natural protein chemistry to create a way to capture, clean up, and release any protein-based sample.
Check out our resources page for more data describing how our technology helps answer new questions about biology.
The ProMTag workflow
All of our protein extraction and cleanup kits include 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 resin
For the generation of peptide samples for mass spectrometry, additional steps include:
- 5) Reduction and alkylation
- 6) Digestion with stabilized and tagged MT-Trypsin
- 7) Capture of MT-Trypsin on ProMTag capture resin
- 8) Elution of clean, digested peptides
For enrichment of membrane proteins, additional steps include:
- 1) Label proteins on the surface of the cell with ProMTag before cell lysis
- 1.1) Wash cells extensively to remove excess ProMTag
- 1.2) Lyse cells with provided cell lysis buffer
- 1.3) Spin out cellular debris and transfer supernatant to ProMTag capture resin
All reagents and supplies for all steps are included in our kits for convenience and reproducibility