Protein quantitation may be accomplished through a number of methods, the most reliable of which include the BCA (bicinchoninic acid) assay and the Bradford assay.Alternative protein quantitation methods include the Lowry assay for protein quantitation and derivitization of N-terminal amines. What is the difference between ELISA and Bradford/BCA protein assay? In this method, a protein can be detected with the help of an antibody. Expert Answer 100% (1 rating) ELISA is an immunological technique. Using standard procedure, the assay is used with samples having protein concentrations between 200 and 1,400 µg/ml (20–140 µg total). Protein concentration can be measured directly, via absorbance at 280 nm in a UV spectrophotometer, or indirectly, using colorimetric methods such as BCA or Bradford assays. The Bradford reagent is an acidified solution of Coomassie G-250; the dye is thus primarily protonated and red. See the answer. Using Microsoft Excel to plot and apply standard curve A protein assay, such the BCA Protein Assay, is an excellent tool for estimating the protein concentration of a sample. The intensity of the colored reaction product is a direct function of protein amount that can be determined by comparing its absorbance value to a standard curve. The Thermo Scientific ™ Pierce BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein. The basis for the Bradford assay is that in order for the Coomassie dye to bind stably to protein, it needs to be doubly protonated. In the Bradford assay, the dye-protein interaction is influenced by the electrostatic interactions of the sulfonate groups with the basic residues, arginine and lysine [4-5].An additional factor is the hydrophobic interactions of the dye with tryptophan, phenylalanine, and tyrosine residues [4,10]. This problem has been solved!
The Bio-Rad protein assay is a simple colorimetric assay for measuring total protein concentration and is based on the Bradford dye-binding method (Bradford 1976). This method combines the well-known reduction of Cu +2 to Cu +1 by protein in an alkaline medium (the Protein composition can bias the results of Bradford and BCA assays. Question: What Is The Difference Between ELISA And Bradford/BCA Protein Assay?
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