UE103C: UbiQuant™ S Kit (Colorimetric)

The UbiQuant™ S assay is a robust tool for assessing the poly-ubiquitination levels of a protein of interest in cell lysate. This platform is based on capturing poly-ubiquitinated proteins on the plate followed by detection of the protein of interest (either native or tagged) using an antibody for the purpose of quantifying the poly-ubiquitinated forms of the protein. Detecting changes in the ubiquitination state of specific proteins in response to external stimuli has been a long and difficult process involving immunoprecipitation followed by Western blot analysis. The UbiQuant™ S kit is a quick, robust, and quantitative alternative to IP/WB analysis. New assay plate developed for colorimetric detection allows for quantitative measurement using absorbance-based plate readers. Colorimetric readouts (650nm, 480nm) are more consistent day-to-day than luminescent-based detection methods, fast, and easy to use. Note: This kit has been optimized and validated for assessing the poly-ubiquitination levels of tagged proteins. Therefore, the end-user must specify the epitope tag in order to receive the appropriate antibody for detection ( myc, HA, FLAG^®, V5, GST.  Unfortunately, His₆ cannot be used for this assay).

UbiQuant™ S Assay Kit Components:

Coated plate:	One pre-coated and blocked 96-well strip plate (Store at -80°C)
Empty strip plate frame:	One empty strip plate frame is provided
Primary Antibody (PA1)	Anti-epitope mAb in PBS Store at 4°C)
Colorimetric reagent (CR):	5.0 mL vial of DR1 reagent (Store at 4°C)
Stopping reagent (SR):	5.0 mL vial of DR2 reagent (Store at 4°C)
Blocking concentrate (BC):	12 mL BC agent (Store at 4°C)
Detection Antibody (DA1):	200X anti-mouse HRP conjugate
Plastic Plate Seals:	Four Plate Seals provided