Millipore Technical Publications |
 | AN- AN2222EN00 | |
The Direct Detect™ Bradford/BCA Comparison Application NoteIR-based Protein Quantitation Surpasses Colorimetric Assay Quantitation and Results are Independent of Detergents, Reducing Agents and Analysis Time | ||
| Lit No: | AN2222EN00 | |
| Year: | 2012 | |
| Abstract Accurate protein quantitation in complex mixtures is crucial for obtaining reproducible biological and biochemical data for both upstream and downstream applications. Colorimetric protein assays are frequently used to determine protein concentration; however, the calculated concentration can vary depending on the sample composition, sequence of the protein, and the time point within the chemical reaction at which the data are collected. Studies involving cell lysates and multiple, diverse proteins (such as multianalyte pathway elucidation) require the development of a rapid, universal protein quantitation method that is compatible with complex samples. Here, we compare infrared (IR)-based protein quantitation using the Direct Detect™ quantitation system to Bradford and bicinchoninic acid (BCA) colorimetric assays and show that IR-based quantitation provides results that match concentrations determined by amino acid analysis, even in the presence of detergent or reducing agent. We also show that, unlike colorimetric assays, the calculated protein concentration obtained from IR-based analysis is unchanged regardless of the time delay between assay and data acquisition. Click the PDF above for the full document. | |
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