How can I prevent degradation / loss of activity in my nuclear protein extract?
To apply a constant workflow, we recommend using the protein extraction kit from Active Motif (#40010 or #40410). In addition, we recommend the following issues for extraction and handling of nuclear proteins: • With the beginning of the extraction procedure, the cells/ tissues and any involved solutions, regents, flasks, tubes, centrifuges etc. must be used on ice. • During extraction, handling, transport, and storage the proteins must be kept on ice. • Avoid centrifugation and “vortex” of final protein extracts. • The extraction has to be performed in the presence of a fresh protease inhibitor mixture (in kits for protein extraction normally included). • Proteins must be stored in 20% glycerol (in kits for protein extraction normally included). • Proteins must be stored at –80°C (-70°C) • Avoid multiple freezing and thawing of final protein extracts. • Store proteins at high concentrations (below 0.75 µg/µl proteins are not stable during storage, even not at –80°C) • Avoid sucrose an
