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Introduction to Biological Sciences

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Added on  2020-05-16

Introduction to Biological Sciences

   Added on 2020-05-16

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Biological Sciences 1BIOLOGICAL SCIENCESBy NameCourseInstructorInstitutionLocationDate
Introduction to Biological Sciences_1
Biological Sciences 2AbstractThe Bradford Protein Assay is a spectroscopic procedure of analysis that helps in the determination of the concentration of proteins in a sample solution. The experiment aims at measuring the concentration of the unknown protein solutions and after that drawing the standardcurve by plotting the 620 nm against a reagent blank. The standards were prepared through the addition of various volumes of BSA stock solution. The absorbances of the unknown samples of protein were determined by the use of a spectrophotometer. A standard curve for the experiment was achieved by plotting 620 nm results the obtained BSA concentration. Linear regression was used in the solution of concentrations of the unknown proteins. Different values were obtained for the various trials that were made, and the average for each trial was calculated.IntroductionA quicker, simpler and safer procedure used in finding the protein concentration in an aqueous solution was developed in the 1970s by Marion Bradford, a biochemist. The procedure has been found to be very efficient thus has become a preference displacing the other methods including Biuret and Folin, which were originally used in the measurement of soluble proteins[CITATION RIa11 \p 159 \l 1033 ]. Because of the disturbance created by the detergents that have been used in the solubilisation process of the membrane-bound proteins, this procedure is not suitable in the form described in this experiment. The original publication of the Bradford Reagent protocol was done in an article Analytical Biochemistry. The protocol underwent refined modification that made it more useful especially for membrane-bound proteins. The modifications were published in the same journal article in the preceding years. The Bradford protein assay is a very convenient method of determining the
Introduction to Biological Sciences_2
Biological Sciences 3concentration of proteins[CITATION Dav128 \p 212 \l 1033 ]. It supplies the dye reagent used in the estimation of the protein concentration at 1x concentrations well as two protein assay standards at prediluted concentrations of seven. The seven prediluted standards are efficiently packaged in screwcap vials of volume 2ml. The packaging reduces and eliminates any form of wasteful sharp ampoules. The packaging also ensures the protein is stable over the product's shelf life. Bradford Protein Assay measures the concentration of proteins in a food sample by adding Coomassie dye to the food sample under acidic conditions. A colour change from brown to blue is noted when proteins in the food samplebind with the Coomassie dye. The concentration or level of the blue colour can be determined using spectrophotometer hence determining the protein concentration in the food sample[CITATION Rob131 \p 251 \l 1033 ].Among the advantage of Bradford Protein Assay are the few steps involved, no need of heating as well as its ability to provide a calorimetric response of greater stability. The response of Bradford Protein Assay method is affected by non-protein sources. It tends to be more nonlinear towards the greatest concentration of protein range[CITATION RIa11 \p 266 \l 1033 ]. The response of this method is also affected by proteins changes with a change in the composition of the protein being tested. Due to these limitations, there is need to standardize the solutions of proteins to achieve results that are more accurate.The amount of light absorbed by a sample is measured by the use of a spectrophotometer. This instrument concentrates a beam through a sample thereby taking measurements of the light intensity that reaches the detector[CITATION Ler12 \p 118 \l 1033 ]. The beam of light is composed of a stream of photons, which are absorbed by an analyte molecule when they come across each
Introduction to Biological Sciences_3

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