The Use of Bradford Assay to Determine the Protein Concentration of Standards and Samples

Abstract Spectrophotometry was performed on three (3) known examines; BSA (bovine blood blood serum albumin), Pepsin, and Lysozyme; and star (1) unknown sample with the use of Bradford assay. This was a chromatic ( disguise changing) order that was used to determine the ducking of proteins in solution. The Bradford assay was a tinge that, when unbound to protein, was in a cationic (positively charged) passing form with and absorbance upper limit of 465 nm. When the dye was bound to protein, it became anionic (negatively charged) and shifted to a blue color with an absorbance maximum of 595 nm. The tot of protein in the sample was determined by the bill of dye that turned blue. The color shift was determined with a spectrophotometer. A standard stoop was constructed before measuring the amount of protein in a sample. A series of five (5) standards contained a ingress ranging from 0 to 1ml of BSA (bovine serum albumin). These standards were assayed and the range of p rotein concentration was connect to their measured absorbance determine. According to the BSA standards analyzed, the density measurements closely related to the concentration BSA present in the standard. For example, at 0.2 mg/ml of BSA an absorption test taken cardinal (2) times averaged out to 0.1485 nm, at 0.4 mg/ml of BSA it was 0.358 nm, at 0.6 mg/ml it was 0.556 nm.

If the values are compared they show that the concentration of BSA is proportional to the absorption. Experimental shift was present from the different testing times and through brusk pipetting technique during dilutions in the Bradford assay. Materials and Metho ds The communications protocol for Experi! ment #1: The use of Bradford assay to determine the Protein assiduity of Standards and Samples was obtained from the BIOL 1F90 Laboratory manual(a) on pages 1-4 (Martin, 2012). Results Table 1: Absorbance after(prenominal) BSA Protein Standard Concentrations are clear to Bradford Reagent SampleBlank12345 Protein (mg/ml)0.00.20.40.60.81.0 Absorbance...If you want to get a full essay, order it on our website: BestEssayCheap.com

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