Quantitation of nucleoprotein complexes by UV absorbance and Bradford assay

Abstract

Despite the importance of studying nucleoprotein complexes, no appropriate method for quantifying them is available. Here, a UV absorbance method using the formula “Cmg/mL = 1.55A280 – 0.76A260” were applied to quantify nucleoprotein complexes. After modification using two paired A260 and A280 values, the UV-derived formula-based method could accurately quantify proteins in nucleoprotein complexes. Otherwise, by taking the target protein as a standard, the Bradford assay can accurately quantify proteins in nucleoprotein complexes without interference by nucleic acids. The above methods were suc-cessfully applied to measure the concentration of MtuP49-CTG complexes of Mycobacterium tuberculo-sis. In conclusion, both the Bradford assay and the UV-derived formula-based method were appropriate for quantifying proteins in nucleoprotein complexes, which may make contributions to explore the interactions between proteins and nucleic acids at the molecular level.