Solution reagent NI™

bovine serum albuminlaboratoryprotein

Protocols.io provides an interactive version of this protocol where you can discover and share optimizations with the research community. A highly sensitive, colorimetric protein assay that overcomes interference of common laboratory agents present in protein solutions and shows minimal protein-to-protein variation. The assay is unaffected by the presence of common laboratory agents, such as reducing agents, chelating agents, detergents, amines, sugars, chaotropes, salts, drugs, antibiotics, cobalt and other common laboratory agents (see tables 1 and 2). The NI™ Protein Assay is composed of two simple steps: Universal Protein Precipitating Agent (UPPA™) is added to the protein solutions to rapidly precipitate total protein. Protein is immobilized by centrifugation and interfering agents in the supernatant are discarded. Protein concentration is assayed by mixing with an alkaline solution containing a known concentration of copper salt; the copper ions bind to the peptide backbone and the assay measures the unbound copper ions (see figure 2). The assay is independent of protein side chains minimizing protein-to-protein variation (see figure 3). The color density is inversely proportional to the amount of protein. The assay is supplied with a traditional bovine serum albumin (BSA) protein standard or a non animal protein standard. Features Linear response 0.5µg-50µg protein Small sample requirement, only 1-50µl Unaffected by non-protein chemicals and agents Protocol time: ~30 minutes Long Shelf Life Applications Estimate protein during protein purification, electrophoresis, cell biology, molecular biology, and other research applications.