IR-based Protein Quantitation Surpasses Colorimetric Assay Quantitation and Results are Independent of Detergents, Reducing Agents and Analysis Time
25 Mar 2012Read this application note to learn about a comparison for infrared (IR)-based protein quantitation using the Direct Detect™ quantitation system to Bradford and bicinchoninic acid (BCA) colorimetric assays. Results show that IR-based quantitation provides results that match concentrations, determined by amino acid analysis, even in the presence of detergent or reducing agent. Also it is shown 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.
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Direct Detect® Spectrometer
MerckIntroducing Direct Detect™, the first infrared (IR)-based biomolecular quantitation system. Just spot your sample, blank, and read! No sample prep, standard curves, messy cuvettes or liquid waste. With inherent amide bond detection, the Direct Detect™ system distinguishes proteins and peptides from interfering sample components, providing more accurate results without the pitfalls of colorimetric assays. Technology: FTIR Spectrometry and Protein Quantitation By measuring amide bonds in protein chains, the system accurately determines an intrinsic component of every protein without relying on amino acid composition, dye binding or redox potential. Accurately analyze components of complex mixtures: proteins, lipids, carbohydrates, and nucleic acids have separable IR spectra. Advantages of FTIR Quantitation: • Independent of amino acid composition • No extinction coefficient required, unlike UV-Vis/A280 methods • Improves speed and accuracy over traditional colorimetric assays • Works in the presence of detergents and reducing agents • Compatible with lysates and membrane preps