To date, detecting changes in the ubiquitylation of specific substrate proteins in response to external stimuli, e.g. stress, cytokine exposure, drug candidate treatment, etc., has been a long, labor- intensive process involving immunoprecipitation followed by gel electrophoresis and Western blot analysis. This method is low through-put, resource intensive, and only semi-quantitative at best. We have developed the UbiQuant™ S kit as a facile, robust, and quantitative alternative to IP/WB analysis. Built on LifeSensors UbiQuant platform, the system captures ubiquitylated proteins in the wells of a microtiter plate and then uses an antibody to the protein of interest (either native or tagged) to quantitate the amount of the protein bound.
Note: If using a tagged-substrate, the user must specify the epitope tag in order to receive the appropriate antibody for detection (e.g. myc, HA, FLAG®, V5, GST, etc. Unfortunately, His6 cannot be used for this assay.)
Determine changes in the level of substrate ubiquitylation as a result of cell treatment.