1. Compute the concentration of the standard solutions by completing Table B.1. Report your answersin 3 decimal places with proper unit. Show one representative calculation only.2. Report the exact values of the linearity constant, slope, and y-intercept of the standard curve.3. Calculate the dilution factor that you will use to calculate for the actual protein concentration.4. Calculate the actual protein concentration in the sample (in mg/mL). 2. The seed extract of Salvia hispanica contains high concentration of protein. In order to quantify theprotein content of Salvia hispanica, 2.5 grams of crude sample was dissolved in 5 mL of tris-HCI buffer.Two more dilutions of the crude sample were made as shown in Table B.2. It was subjected then toBradford assay to determine its concentration against an external calibration standard as shown inTable B.1. Three-mL each of bovine serum albumin (BSA) standard solutions were prepared from astock solution with concentration of 20 mg/mL. The relevant information is shown below:Table B.1. Absorbance readings of the BSA standard solutions.Initial volume of BSA stocksolution, mL0.000.150.300.450.600.75Concentration,mg/mL(1)(2)(3)(4)(5)(6)1 in 2 dilution1 in 5 dilutionAbsorbance at540 nmTable B.2. Absorbance readings of the crude extract:SampleUndiluted0.0000.1240.2010.2980.3630.434Absorbance at 540 nm0.5480.4380.244

Bradford assay is a method to estimate the protein concentration in the given sample. It is based on…

Answered: 1. Compute the concentration of the…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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1. In the preparation of the standard curve for protein analysis, 50 mnBSA (bovine serum albumin) dissolved in H2O to a final volume of 5mL was used as stock solution. What is the weight of BSA in 0.1 mL ofstock solution? In 0.2 mL? 2. The above mentioned aliquots (#9) which were diluted with enoughwater to a final volume of 1 mL were assayed colorimetrically andyielded the following absorbance readings:mL stock mL H2O Absorbance0.2 0.8 0.1000.4 0.6 0.2000.6 0.4 0.3000.8 0.2 0.4001.0 0.0 0.500 i. Tabulate mg BSA vs. absorbance readingii. Draw the curve where mg BSA is on x-axis and absorbance ony-axis (use graphing paper).iii. Calculate the slope of the curveiv. If a sample solution has an absorbance of 0.332, determinemg protein of the sample.
1. In the preparation of the standard curve for protein analysis, 50 mnBSA (bovine serum albumin) dissolved in H2O to a final volume of 5mL was used as stock solution. What is the weight of BSA in 0.1 mL ofstock solution? In 0.2 mL? The mentioned aliquots (#9) which were diluted with enough water to a final volume of 1 mL were assayed colorimetrically and yielded the following absorbance readings: mL stock mL H2O Absorbance0.2 0.8 0.1000.4 0.6 0.2000.6 0.4 0.3000.8 0.2 0.4001.0 0.0 0.500 i. Tabulate mg BSA vs. absorbance readingii. Draw the curve where mg BSA is on x-axis and absorbance ony-axis (use graphing paper).iii. Calculate the slope of the curveiv. If a sample solution has an absorbance of 0.332, determinemg protein of the sample.
Bradford Assay Fill in the average A595 and A595 of sample minus A595 of blank. Show some sample computation. Concentration (micrograms/ml) A595 - trial 1 A595 - trial 2 A595 - trial 3 average A595 A595 of sample minus A595 of blank 0 1.501 1.446 1.447 2 1.624 1.558 1.559 5 1.731 1.749 1.712 10 1.901 1.838 1.892 15 2.161 2.108 2.228 18 2.231 2.277 2.319
Match the types of data to the example. Precise measurements Assay standard Positive control Quantitative results Negative control Qualitative results Standard curve…
Various concentrations of recombinant human insulin were prepared for use standards for an HPLC method. To verify the prepared concentrations, the samples were analyzed by measuring the absorbance at 280 nm in a short path length (5 mm) cuvette. The molar absorption coefficient for human insulin is approximately 5.875 x 10³ M-¹cm-¹. a. Calculate the extinction coefficient in mL mg-¹cm-¹. b. Calculate the concentrations of the following human insulin standards if the measured absorbances and dilutions used are: Standard 1 Standard 2 Abs. (at 280 nm) of Diluted Sample 0.305 0.685 Dilution 145.0 µL sample, 25.0 μL buffer 130.0 µL sample, 40.0 μL buffer
A series of bovine serum albumin (BSA) was prepared and 1 mL of each solution was subjected to a Bradford assay. The increase in absorbance at 595 nm relative to the concentration of protein was plotted as shown below. Save 1.0- 0.8- 0.6- 0.4- 0.2- 0.04 0.0 0.5 1.0 2.0 2.5 1.5 BSA concentration (mg/mL) a) Using the graph, calculate the original concentration of protein present in a mixture of haemoglobin and methylene blue (diluted 1:50), which gave an absorbance of 0.20 at a wavelength of 595 nm using the Bradford assay. Show all workings. Absorbance (O.D.)
7. A total of 7.21 g of dry mixture with a protein content of 89.2% (w/w) determined by Kjeldahl analysis was dissolved in 88.54 g of sterile water and centrifuged to purify the protein in the mixture. After centrifugation, a total of 5.95 g of precipitated pellet was formed, and the protein content in pellet was determined as 72.2% by spectrophotometric Lowry analysis. Assume that pellet is not a hygroscopic material and does not absorb any water. Determine the following: a. the protein concentration in the supernatant layer after centrifugation in % w/w b. the nature of amino acids in the protein structure. What's the possible explanation?
1. What is the function of each step or reagent used in the isolation of casein from milk? Reagents: a. Warm distilled water b. Vinegar c. Ethanol 2. What are the three general steps involved in the isolation of proteins? Discuss or briefly describe the purpose of each step. 3. Suggest a simple procedure (or test) to verify if the final product you obtained is indeed casein.
2. The following data were generated after a set of bovine serum albumin standards were measured via the BCA assay. Table 1. Absorbance measurements at 562 nm for BSA standards measured in triplicate in a BCA Assay. Standards A B C D E F G H I Conc (ug/ml) 2000 1500 1000 750 500 250 125 25 0 Replicate 1 Abs @ 562 nm 2.381 2.039 1.505 1.143 0.819 0.454 0.245 0.056 0 Replicate 2 Abs @ 562 nm 2.456 2.052 1.489 1.156 0.823 0.46 0.272 0.049 0 Replicate 3 Abs @ 562 nm 2.43 2.05 1.47 1.17 0.81 0.48 0.255 0.053 0.00 Use Excel or your graphing program of choice to create a standard curve using the data from table 1. Consider the best way to graph this data. Make sure the type of graph that you select will allow you to add a line of best fit. Display the equation for the line of best fit directly on the graph. Hint... you'll want to think about how to treat the triplicate values when making the graph. Also consider the type of graph that should be selected to allow for addition of the line of…
During incubation, prepare dilutions of the standard antiserum which constitutes the standard curve for the assay (concentration in ng/mL). Add 500 µL of PBS-milk to the supplied microtube containing 500 µL of standard antiserum to obtain Standard 1 [500 ng/mL]. Identify seven microtubes for standards 2 to 7 and place 500 µL of PBS-milk in each. Calculate how much of antiserum is used in each standard ?
Calculation for a neutral red assay please include steps since it is essential for the report and it is marked.
1. Following the modified protocol for the isolation of Escherichia coli bacteriophage of Encabo (2018) presented below, compute for the pfu/ml of the chloroform-treated lysate. 1ml 1ml 1ml 1ml 1ml | | ||| 9ml 9ml 9ml 9ml 9ml Chloroform-treated lysate 101 102 103 104 105 0.1 ml 0.1 ml_0.1 ml Empty sterile tubes ||| + 0.5 ml E. coli Incubate inside ref for 15-20min + 0.1 ml lysate-E.coli mix Molten soft agar overlay O00000 Bottom agar Number of plaque-forming units (pfu) Plate A Dilution Plate B 103 104 105 254 265 132 11 110 23 average number of plaques x dilution factor volume plated pfu ml

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