FAQ
How do I minimize protein degradation in sample preparation for electrophoresis?
During sample preparation (e.g., cell fractionation) endogenous proteases are released which may degrade sample proteins. To prevent artifacts caused by proteolysis, use protease inhibitors during sample preparation and protein isolation.
For SDS-PAGE, deactivation of endogenous proteases may require mixing the sample with SDS and heating to 70 °C - 100 °C for 3 minutes. This cannot be done for native electrophoresis samples. To minimize proteolysis the run should be performed at 4 °C.
What samples have been labeled using the Ettan DIGE technology?
A. Mammalian cell lines, mammalian tissues, mammalian biopsy, bacteria, yeast, plant, Drosophila, and biological fluids such as plasma and serum.
Do the bound CyDye DIGE fluors block protease activity?
We have no evidence of this, but it is likely that after the CyDye DIGE fluor has covalently attached to a lysine amino acid that a single trypsin digest site will be lost.