Proteolysis of protein mixtures in sera or gels is a fundamental step in "Bottom Up" workflows for protein characterization. Lipids and detergents are harmful downstream of this point so it is important to remove them before or during digestion, else one must fractionate the proteins using denaturing PAGE. To be consistently successful, it is critical to pay careful attention to many parameters such as turbidity, viscosity and surface tension across all of the steps. This is why we prefer to perform digestions in the core, especially for new projects, where immediate troubleshooting is relatively convenient. If you will execute the digests, please look over procedures and practices that have been successful for us. We have utilized a variety of sources to develop these procedures.
