Suppose that you are tasked to determine the protein concentration of an unknown protein solution via Bradford assay. You prepared six solutions of bovine serumalbumin (BSA) with different concentrations. The initial concentration of the BSA stock solution is 7.50 mg/mL. Approximately 200 µL of Bradford Reagent was added toeach of these solutions and the absorbance at 595 nm was taken after 5 minutes. See the table below for data on the standard solutions.Standard #A595BSA conc (mg/mL)0.0000.15821.1250.2912.2500.37243.3750.50354.50065.6250.675Determine the protein concentration, in mg/mL, of the unknown solution if its absorbance at 595 nm was 0.248.Note: Final answer format must be x.xx (two decimal places). Round off only in the final answer. Do not round off in the middle of calculation.

The concentration of unknown protein solution can be determined by plotting a graph between…

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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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Similar questions

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The bacteria that you are growing to express biochemisfunase are resistant to ampicillin, so you keep ampicillin in your media to prevent contamination from other bacterial strains. You find a tube of ampicillin in the lab freezer that says “2000x AMP” on the lid, which indicates that it is 2000 times more concentrated than what you need (i.e., a concentration of 1x ampicillin in your media). How much of the 2000x ampicillin stock would you add to your 1.0-L culture to get a final concentration of 1x ampicillin?
The data in the table are used to create a calibration curve for the determination of RNA from ts absorbance at 260 nm. Use a spreadsheet and the least-squares method to determine the slope and y-intercept of the best straight line fit to the calibration curve. Do not subtract the blank reading when creating the calibration curve. m= b = RNA (µg) 0.00 10.36 20.72 31.08 41.44 Absorbance at 260 nm 0.268 0.497 0.795 1.131 1.373 µg Using the slope and intercept of the calibration curve, determine the quantity of RNA in a sample that gives an absorbance of 0.699. -1
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Standard curves You are setting up a standard curve for lysozyme. You have a stock vial containing 2 mg/ml of lysozyme and you have 2 ml in the vial. You will want to have a minimum of 1 ml of each concentration for the spectrophotometer. How would you proceed? (Saline is added to make the various concentrations). Concentration Am't stock lysozyme or am't of a previous dilution (note which) Amount saline Total volume wanted 1.00 mg/ml 0.75 mg/ml 0.50 mg/ml 0.375 mg/ml
The data in the table are used to create a calibration curve for the determination of RNA from ts absorbance at 260 nm. Use a spreadsheet and the least-squares method to determine the slope and y-intercept of the best straight line fit to the calibration curve. Do not subtract the blank reading when creating the calibration curve. m = b = RNA (µg) 0.00 10.36 20.72 31.08 41.44 Absorbance at 260 nm 0.268 0.497 0.795 1.131 1.373 Hig Using the slope and intercept of the calibration curve, determine the quantity of RNA in a sample that gives an absorbance of 0.699.
A protein gives a single band on SDS gel electrophoresis, as shown in lanes 1 and 2 below. There is little, if any, effect from adding β-mercaptoethanol(BME) to the sample; if anything, the protein runs a little bit slower. When treated with the proteolytic enzyme thrombin and electrophoresis in the absence of BME, the protein migrates a bit more rapidly (lane 3). But if BME is present, two much more rapidly migrating bands are found (lane 4). Explain these results in terms of a model for the protein.
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You are given an equimolar (0.10 mM) mixture of Ubiquitin protein that is free/pure of any other macromolecules (e.g. nucleic acids) in a pH 7.0 phosphate buffer How can you purify Ubiquitin? Propose a 2-step minimum plan to purify their protein. Basic rules: Affinity chromatography (e.g. His Tag purification) is not an allowed step because these proteins are in their native state (i.e. – do not have a polyhistidine tag). Also, you are not allowed to rely solely on the color of certain proteins (e.g. cytochrome C and GFP) in your characterization/proposal. You are allowed to buffer exchange (i.e. – switch buffers) but try to keep your pH’s in a reasonable range (5.5 – 8.5) or you risk denaturing your protein! Assume you can only separate proteins under 50.0 kDa with a 5.0 kDa difference using SEC. Otherwise, assume proteins greater than 50.0 kDa are unable to be separated. Assume proteins with the same charge (positive or negative) are not easily separated using IEC. Assume all…
Prepare the following LB media with antibiotic added. A 100ml of LB media with 25 micro/ml of Amp and 100micro/ml of Kan final concentration. A 100ml of LB is provided and amp of Kan stock at 100 mg/ml and 50 mg/ml are also provided. Find how much each antibiotic stock solution is needed to add 100ml of LB to reach desired antibiotic concentration.

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