Principle of Assay
This kit is similar to the traditional BCA protein quantitation method, but employs a completely new and specific chelator different from BCA, resulting in a rapid, stable, and sensitive determination of protein concentration. The principle of the method is that the protein reduces copper ion (Cu2+) to cuprous ion (Cu+) in basic conditions, and the generated Cu+ forms an orange yellow water-soluble complex with chelates and has a strong absorption peak at 480 nm, the absorbance value is proportional to the content of protein in the sample, and the protein concentration can be determined according to the absorbance value. The chelators in this kit can sensitively and specifically bind to Cu+ and complete the color reaction with only 5 min incubation at room temperature. In response to the pain points of the traditional BCA assay method such as long time consuming (up to 30-90 min), unstable results, and low sensitivity, this kit has been developed to effectively solve the problems in protein determination, which has the following worry-free advantages: (1) Fast and time-saving: 5 min room temperature color development reaction; (2) Wide measurement range: wide linear working range, 1-2000 µg/mL protein concentration; (3) Accurate absorbance: the colorimetric method is used to have the best absorbance at 480 nm; (4) High sensitivity: detection of protein concentration as low as 1 µ/mL; (5) Sample volume to be measured: 1-20 µL. The protein concentration determined by method is not affected by chemical substances in most samples, and can be compatible with up to 5% SDS, 5% Triton X-100, 5% Tween 20, 60, and 80 in the sample. However, the method is affected by chelating agents and slightly higher concentrations of reducing agents. It is necessary to ensure that EDTA is less than 10 mM, no dithiothreitol (DTT) and beta-mercaptoethanol.
