Bradford assay was used to determine bovine serum albumin (BSA) concentrations. The method works by binding Coomassie Brilliant Blue dye to protein standards and unknown, leading to a shift in the absorbance maximum of the dye. A stock solution of BSA is available (10 mg/ml) from which the protein standards were prepared. Bradford reagent, 290 ul, was pipetted into a transparent 96-well microplate. 10 ul of the protein dilution was added followed by mixing in the wells. After 5 min of incubation at room temperature, the plate was read in spectrum mode with the absorbance spectrometer of the microplate reader. The spectrum of the dye not bound to protein was also determined. Create a standard curve or regression equation that related the protein concentration and absorbance. An unknown BSA sample was prepared the same way as the standard and the OD was 0.35. determine the original concentration of the protein solution. The measurements can be seen below. Protein Concentration Average Absorbance 0.0625 0.02 0.125 0.03 0.25 0.05 0.5 0.09 1 0.22

Bradford assay was used to determine bovine serum albumin (BSA) concentrations. The method works by binding Coomassie Brilliant Blue dye to protein standards and unknown, leading to a shift in the absorbance maximum of the dye. A stock solution of BSA is available (10 mg/ml) from which the protein standards were prepared. Bradford reagent, 290 ul, was pipetted into a transparent 96-well microplate. 10 ul of the protein dilution was added followed by mixing in the wells. After 5 min of incubation at room temperature, the plate was read in spectrum mode with the absorbance spectrometer of the microplate reader. The spectrum of the dye not bound to protein was also determined. Create a standard curve or regression equation that related the protein concentration and absorbance. An unknown BSA sample was prepared the same way as the standard and the OD was 0.35. determine the original concentration of the protein solution. The measurements can be seen below. Protein Concentration Average Absorbance 0.0625 0.02 0.125 0.03 0.25 0.05 0.5 0.09 1 0.22

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

Chapter9: Atomic Absorption And Atomic Fluorescence Spectrometry

Section: Chapter Questions

Problem 9.22QAP: The chromium in an aqueous sample was determined by pipetting 10.0 ml. of the unknown into each of...

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The chromium in an aqueous sample was determined by pipetting 10.0 ml. of the unknown into each of five 50.0-mL volumetric flasks. Various volumes of a standard containing 12.2 ppm Cr were added to the flasks, following which the solutions were diluted to volume. Unknown,mLStandard, mLAbsorbancc 10.00.00.201 10.010.0 0.292 10.020.0 0.378 10.030.0 0.467 10.040.0 0.554 (a) Plot the data using a spreadsheet. (b) Determine an equation for the relationship between absorbance and volume of standard. (c) Calculate the statistics for the least-squares relationship in (b). (d) I)ctcrmine the conccnt ration oÍCr in ppm in the sample. (e) Find the standard deviation of the result in (d).
In a Bradford assay, 17 pl of a protein isolate sample was diluted by adding 24 pl 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 0,384 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 ug/mL The concentration and the corresponding absorbance reading at 595 nm of the protein standard solutions are summarized in the table below: Standard soln # 3 6 conc of BSA Kug/mL) 12.00 24.00 36.00 48.00 60.00 Absorbance lat 595 nm 0.000 0.156 0.249 0.328 0.411 0.528 zNote: Final answer format must be x.xox (three decimal places) Round off only in the final answer. Do not round off in the middle of the calculation
The standard curve of a biuret assay gave the equation: y=0.4193x. The absorbance for Unknown A = 0.098 and Unknown B = 0.325. Are you equally confident in the accuracy of the protein %D concentrations you would calculate for samples Unknown A and Unknown B? Briefly explain your answer.
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Protein ABC is a heme-containing protein responsible in electron transfer in microalgae. The target protein was extracted using ammonium sulfate fractional precipitation and quantified using Bradford assay. A set of solutions with increasing concentration was prepared using 15.00 mg/mL analogous protein standard [refer to succeeding table]. Each solution was treated with 80 μL of Bradford reagent and diluted to 1.500mL. The absorbance at 595 nm of each solution was obtained and presented below. Volume of protein std, μL A595 0 0.0002 10 0.0007 25 0.0075 50 0.0165 100 0.0356 250 0.0930 500 0.1950 750 0.3010 1000 0.4175 Report the equation of the calibration curve and proof of linearity (R2 value).
A third spectrophotometric method for the quantitative analysis of Pb2+ in blood uses Cu2+ as an internal standard. A standard containing 1.75 ppb Pb2+ and 2.25 ppb Cu2+ yields a ratio of (SA/SIS)std of 2.37. A sample of blood is spiked with the same concentration of Cu2+, giving a signal ratio, (SA/SIS)samp, of 1.80. Determine the concentration of Pb2+ in the sample of blood.
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).
A 5.00-mL sample of blood was treated with trichloroacetic acid to precipitate proteins. After centrifugation, the resulting solution was brought to pH 3 and extracted with two 5-mL portions of methyl isobutyl ketone containing the lead-complexing agent APCD. The extract was aspirated directly into an air/acetylene flame and yielded an absorbance of 0.527 at 283.3 nm. Five-milliliter aliquots of standard solutions containing 0.400 and 0.600 ppm of lead were treated in the same way and yielded absorbances of 0.396 and 0.599. Find the concentration of lead in the sample in ppm assuming that Beer’s law is followed.
CPU/RAM ProctorU e Proctor e Proctoru o.com/collab/ui/session/join/43918c0bf9cb421ca7b98b0075f5f239 Firefox Ex SPECTROSCOPY 1. If the concentration of solute in a solution is high, the Absorbance will be high or low and why? 2. Was the spectroscopy experiment qualitative or quantitative analysis? 3. If we have 12M HCI and we need 3 L of 1M HCI, how much 12 M should we use? 4. What does Molarity mean? 5. When must safety goggles be worn? 6. If you make a mistake recording data, what must you do? spec hw.pptx (2/3) ..% An
A company is producing a powder which can be reconstituted as a healthy drink for the elderly. Analysis of the powder for calcium involved using the following procedure. 4 g of powder was reconstituted to 150 ml with water. Thirty ml of 24 % TCA was added to 30 ml of this solution. The mixture was allowed to stand for 15 mins and then filtered. The filtrate was analysed for calcium using atomic absorption spectroscopy. From the standard curve it was found that the reading for the analysed sample was 24mg/litre. Calculate the concentration of calcium in the powder (mg/kg).
The Bradford method described in the Background section was used to determine protein concentrations of known and unknown samples The following results was obtained: Table 1: Absorbance at 595nm obtained for different concentrations of protein standards using the Bradford method Protein (mg/ml) 0.00 0.125 0.25 0.50 0.75 1.00 1.50 2.00 Unknown Abs. BSA 0 0.130 0.350 0.620 0.870 1.120 1.400 1.600 0.635 Abs. BIg 0 0.080 0.230 0.460 0.520 0.410 0.510 0.700 Calculate the concentration of the unknown protein in mg/ml
Exactly 5.00 mL aliquots of a solution containing analyte X were transferred into 50.00-mL volumetric flasks and the pH of the solution is adjusted to 9.0. The following volumes of a standard solution containing 2.00 µg/mL of X were then added into each flask and the mixture was diluted to volume: 0.000, 0.500, 1.00, 1.50 and 2.00 mL. The fluorescence of each of these solutions was measured with a fluorometer, and the following values were obtained: 3.26, 4.80, 6.42, 8.02 and 9.56, respectively. ii. Using relevant functions in Excel, derive a least-squares equation for the data, and use the parameters of this equation to find the concentration of the phenobarbital in the unknown solution.