Amplite(TM) Rapid Colorimetric Maleimide Quantitation Kit

Ex: 780 nm. A variety of crosslinking reagents with a maleimide group are widely used for crosslinking proteins to proteins, or proteins to other biomolecules. A key challenge in maleimide crosslinking technology has been the quantitation of maleimide linked to a single protein or antibody. Maleimides can be directly assayed spectrophotometrically at 302 nm. However, the small extinction coefficient of 620 M-1cm-1 renders this assay insensitive, and the assay is further complicated by the protein absorbance at the same wavelength. AAT Bioquest's AmpliteT Rapid Colorimetric Maleimide Quantitation Kit provides a rapid and accurate method to quantify maleimide using our proprietary maleimide sensor Maleimide BlueT with the maximum absorbance at ~780nm. The principle of this assay is that Maleimide BlueT reacts with the maleimide-linked sample, and the resulted product is run through a single spin column to remove the excess sensor. The absorption spectrum of the purified product is measured, and the amount of maleimide to protein ratio can be calculated from the absorbance ratio of 780 nm and 280 nm. This AmpliteT Rapid Maleimide Quantitation kit can be performed in a traditional cuvette, NanoDropT Spectrophotometer or a convenient 96-well absorbance plate reader with a UV-transparent plate.