3. Incubate the solution at room temperature for 5 min.4.Read the absorbance at 595nm of the blank and test solutions.Calculate the protein concentration (in mg/ml) of an unknown sample with an absorbancy value of 0.996 using the standard curve below.1.51.41.31.21.11.00.90.80.70.60.50.40.30.20.10.00.00.10.20.30.40.50.60.70.80.91.01.11.21.31.41.51.6BSA concentration (mg/ml)wuseS Áɔuequosqy QUESTION 3An unknown protein was determined using the following method:Bradford methodReagentsStock Bradford reagent: Dissolve 100 mg Coomassie Brilliant Blue G250 in a mixture consisting of 100ml of 85% phosphoric acid, 50ml of 95% ethanol and 50ml 1M NaOH.Store at 4°C until precipitation occurs, at which point it is discarded.Working Bradford reagent: Prepare fresh by diluting 10ml of stock Bradford reagent to 250ml with distilled water.Stock bovine serum albumin (BSA) solution (10mg/ml): Dissolve 0.2g BSA in 20ml distilled water.Working BSA concentration range: 0.5 – 1.25 mg/mlMethod1. Prepare the following test tubes:Table 1: Preparation of test tubes for the Bradford methodBlankTube 1Tube 2Tube 3Tube 4Tube 5UnknownDistilled H201.0 ml0.9ml0.9ml0.9ml0.9ml0.9ml0.8ml0.25 mg/ml BSA0.1 ml0.5 mg/ml BSA0.1 ml0.75 mg/ml BSA0.1 ml1.0 mg/ml BSA0.1 ml1.25 mg/ml BSA0.1mlUnknown0.2mlproteinWorking4.0ml4.0ml4.0ml4.0ml4.0ml4.0ml4.0mlBradford2.After each addition of the working Bradford reagent, thoroughly mix the resultant solution.

Bradford method of protein estimation is quick and simple method of protein estimation using…

Answered: Calculate the protein concentration (in…

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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To prepare a gel sample, you want to load 50 ng total of protein/well. You have added 200 μL of protein in 800 µL reagent and measured your sample by Bradford A595 to be 0.7 mg/mL - your dilution is unaccounted for at this point. Assuming a total final volume of 20 μL, what volume of protein sample, buffer and 4X SDS PAGE Loading Dye needs to be mixed to create a final sample of 50 ng protein in 20 μL?
The following stock solutions are available to make a protein extraction buffer: 100% Nonidet P-40, 1 M Tris-Cl, and 0.5 M EDTA. What quantity of the original stocks will be needed to make 250 ml of buffer with the following final concentrations: 0.5% Nonidet, 150 mM Tris-Cl, and 10 mM EDTA?
Protein concentration standards 0.6 0.5 0.4 0.3 0.2 0.1 0.5 1.5 2.5 Albumin concentration (mg/mL) A standard curve of known concentrations of protein were reacted with the Biuret reagent. A linear curve fit is presented above. • Use the graph to estimate the concentration of an unknown sample with an A550 value of 0.416 (A rough estimate is fine. You do not need a calculator for this problem) • Use the graph to estimate the concentration of an unknown sample with an A550 value of 0.233 • Which instrument did we use to measure the intensity of the color of proteins that reacted with Biuret reagent?
Size Size Crude Anion Cation 2) Exclusion Exclusion Lysate Exchange Exchange 1 2 Total Protein Concentration 15.2 6.6 2 3.75 4.7 (mg/mL) Final Sample Volume 60 30 20 4 3 (mL) Enzyme Specific Activity 0.43 1.7 7.25 18.3 12.5 (units/mgprotein) Based on your protein purification sample data, in which of the purification steps in your protocol did you effectively purify your enzyme? Select all that apply. a) anion exchange correct b) cation exchange correct c) size exclusion 1 correct d) size exclusion 2
Using DEAE-cellulose as ion exchange resin, indicate the starting and ending pH for the narrowest experimental pH range used to separate an amino acid mixture consisting of Gln, Leu and Lys Starting pH: _____ Ending pH: _____
N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) buffer at pH 7 with 400 mM NaCl. A A purified protein is in a Hepes dialysis membrane tube holds a 5.0 mL sample of the protein solution. The sample tube floats in a beaker containing 0.50 L of the same Hepes buffer, but with 0 mM NaCl, for dialysis. Small molecules and ions (such as Na*, CI", and Hepes) can to diffuse across the dialysis membrane, but the protein cannot. Assume there are no sample volume changes during the dialysis. Calculate the final concentration of NaCl in the protein sample once the dialysis has come to equilibrium. Calculate the final NaCl concentration in the 5.0 mL protein sample after dialysis in 300 mL of the same Hepes buffer, with 0 mM NaCl, twice in succession. [NaCl] after a single dialysis: [NaCl] after a double dialysis: mM mM
10 tab of acetaminophen 500 mg molar absorptivity= 14520 @ 243nm. The total weight of the 10tab is 5278 mg. I take a sample equivalent to 100mg of tablet and I diluted it in 100mL, I took an aliquot of 10mL and I diluted it in 250mL, of these I took 25mL and I diluted them in 250mL. Calculate the weight per tablet. Absorbance = 0.944
Given the following information, calculate the total activity in the undiluted protein sample. Activity of 1 ml of diluted sample = 0.5 Total volume of sample = 5 ml Dilution factor = 10 25 50.5 250 2.5
Laemmli buffer, also referred to as SDS-reducing buffer, is added to each of the samples. It is a blue solution and the recipe for 2X Laemmli buffer is: 0.125 M Tris HCl, pH 6.8, 20% glycerol, 4% SDS and 10% β-mercaptoethanol (βME). What is the purpose of each of these ingredients?
COLOR REACTION OF PROTEINS Question Guide: 1. Write the positive result, the chemical group responsible for the positive and the importance of each test. Test Positive Result Chemical Group Biuret Ninhydrin Xanthoprotei Millon's Hopkins-Cole Lead Acetate Pauly Bromine Water Sakaguchi
can you show how to calculate the amount of two components for the required volume of a buffer with the receipes? Prepare enough 10X stock solution § Prepare 1LProportions§ 0.25M tris§ 1.92M glycine§ pH 8.3§ 1% (w/v) SDSInstructions to prepare 100mL of 10X running buffer1. Weigh 3.02g of Tris base and 14.42g of glycine into ~90mL of ddH2O in a clean beaker and stir todissolve.2. Check the pH with a calibrated pH metre once dissolved.§ The pH of the buffer should be 8.3. pH adjustment is not required if between 8.1-8.5.3. Add 5mL 20% (w/v) SDS stock solution (a liquid) to make a 1% (w/v) solution.4. Top the volume to 100mL by adding 5mL ddH2O from a graduated cylinder.
How many mL of the 1 M glucose stock solution do you need to prepare 100 mL of a 1 mM glucose solution?

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