You obtained the following raw data when setting up a Bradford standard curve:BSA (mg/ml)Absorbancy595nm00.15510.28720.42030.52340.65150.75660.87870.91880.9549100.9820.990After blanking against a bradford-dH2O sample, the protein concentration of an unknown sample was determined using the same method andan absorbancy of 0.566 was obtained.Set up a standard curve, excluding outliers (experimental and statistical) and determine the protein concentration in the unknown sample inmg/ml (up to 3 decimal places).

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Answered: You obtained the following raw data…

Principles of Instrumental Analysis

7th Edition

ISBN:9781305577213

Author:Douglas A. Skoog, F. James Holler, Stanley R. Crouch

Publisher:Douglas A. Skoog, F. James Holler, Stanley R. Crouch

Chapter27: Gas Chromatography

Section: Chapter Questions

Problem 27.19QAP

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In a Bradford assay, 11 µl of a protein isolate sample was diluted by adding 21 µl of water prior to the addition 2.0 mL Bradford reagent. Calculate the protein concentration (in mg/mL) of the original protein isolate sample whose absorbance reading at 595 nm is 0.304 after dilution. The protein standard solutions used for the calibration curve of the Bradford assay was constructed using bovine serum albumin (BSA) stock solution whose concentration is 200 µg/mL. The concentration and the corresponding absorbance reading at 595 nm of the protein standard solutions are summarized in the table below: Standard soln # 1 2 13 4 6 conc of BSA 12.00 24.00 36.00 48.00 60.00 (pg/mL) Absorbance at 595 nm 0.000 0.156 0.249 0.328 0.411 0.528 zNote: Final answer format must be x.xxx (three decimal places). Round off only in the final answer. Do not round off in the middle of the calculation.
You obtained the following raw data when setting up a Bradford standard curve: BSA (mg/ml) Absorbancy 595nm 0 0.225 1 0.310 2 0.420 3 0.510 4 0.610 5 0.720 6 0.810 7 0.915 8 0.950 9 0.980 10 0.990 After blanking against a bradford-dH2O sample, the protein concentration of an unknown sample was determined using the same method and an absorbancy of 0.523 was obtained. Set up a standard curve, excluding outliers (experimental and statistical) and determine the protein concentration in the unknown sample in mg / ml (up to 3 significant figures).
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Plot a professional quality standard curve (absorbance vs. protein amount in µg) with an appropriate title and caption underneath the plot from the data give above. The caption should include the best-fit data.
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Aliquots of a 0.5 mg/mL standard of BSA were used to construct a standard curve for the Bradford protein assay. The tubes contained the following amounts of the BSA solution: 0, 20, 40, 60, 80, 100 μL. Each sample then had water added to make the final volume 100 μL. The corresponding absorbencies after adding Bradford reagent were the following: 0, 0.05, 0.09, 0.14, 0.19, and 0.22. If you took 20 μL of an unknown and added 80 μL of water, mixed, took 10 μL of the mixture and 90 μL of water and added the Bradford reagent and saw an absorbance of 0.08, what was the protein concentration of the undiluted unknown to one decimal place?

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