Estimation of Protein Concentration Practical
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Questions and Answers

What is the main objective of the practical on estimation of protein concentration?

  • To detect kidney malfunction through proteinuria
  • To estimate the relative number of peptide bonds in different proteins (correct)
  • To analyze dietary requirements in foodstuffs
  • To determine the amount of an enzyme in a solution

    • Why is accurate protein concentration determination important in biochemical molecular research?

  • For all aspects of biochemical molecular research (correct)
  • To determine the amount of an enzyme in a solution
  • To detect kidney malfunction
  • To analyze dietary requirements

    • What is the primary method of protein determination used in the practical?

  • The Biuret reaction (correct)
  • Determination of enzyme amount in a solution
  • Analysis of dietary requirements
  • Detection of kidney malfunction

    • How many principal methods of protein determination are there?

    <p>5</p> Signup and view all the answers

    What is the weight of albumin used as a 'standard' protein in the practical?

    <p>5 mg</p> Signup and view all the answers

    Why is the completion of the test on Blackboard important?

    <p>To prepare for the other practicals in the module</p> Signup and view all the answers

    What is proteinuria used for in biological samples?

    <p>To detect kidney malfunction</p> Signup and view all the answers

    What is the purpose of estimating protein content in plasma derived from blood?

    <p>To determine the protein content of plasma</p> Signup and view all the answers

    What is the minimum number of peptide bonds required for the Biuret reaction?

    <p>2</p> Signup and view all the answers

    What is the significance of peptide bonds in protein determination?

    <p>To estimate the relative number of peptide bonds</p> Signup and view all the answers

    Why is this practical not assessed?

    <p>Because it is a preparatory practical for other assessed practicals</p> Signup and view all the answers

    What is the primary limitation of protein assays that depend on the reactions of individual amino acids?

    <p>They are influenced by the variation in amino acid composition of proteins</p> Signup and view all the answers

    What is the purpose of adding Folin Ciocalteau's reagent in the Lowry method?

    <p>To enhance the Biuret reaction</p> Signup and view all the answers

    What is the general percentage of nitrogen in proteins?

    <p>16%</p> Signup and view all the answers

    Why is the nitrogen content method frequently used for solid samples?

    <p>Because it can cope with insoluble samples</p> Signup and view all the answers

    What is the wavelength of ultraviolet absorption used to estimate protein concentration?

    <p>280 nm</p> Signup and view all the answers

    What is the primary advantage of the dye-binding method?

    <p>It does not react with free amino acids</p> Signup and view all the answers

    What is the purpose of wearing gloves and safety spectacles in the laboratory?

    <p>All of the above</p> Signup and view all the answers

    What should you do in case of skin or eye contamination?

    <p>Rinse with lots of water and inform a demonstrator</p> Signup and view all the answers

    What is the purpose of the Accident Book in the laboratory?

    <p>To report accidents and near-misses</p> Signup and view all the answers

    Why is it important to dispose of waste chemicals properly at the end of the practical?

    <p>To prevent contamination</p> Signup and view all the answers

    What is the primary purpose of using a new pipette tip for each separate solution?

    <p>To prevent contamination of the Biuret reagent</p> Signup and view all the answers

    What is the total volume in each tube after adding 4 ml of Biuret reagent?

    <p>5.0 ml</p> Signup and view all the answers

    What is the purpose of leaving the mixture for 20 minutes before measuring the absorbance?

    <p>To allow the Biuret reagent to react with the proteins</p> Signup and view all the answers

    What is the unit of measurement for the protein concentration on the x-axis of the graph?

    <p>mg per ml</p> Signup and view all the answers

    What is the purpose of adjusting to zero absorbance using the contents of Tube 1 as the blank and reference solution?

    <p>To eliminate the background interference in the measurements</p> Signup and view all the answers

    What is the equation of the line of best fit used for?

    <p>To estimate the concentrations of protein in the casein, gelatin, and plasma solutions</p> Signup and view all the answers

    What is the purpose of using the contents of Tube 1 as the blank and reference solution?

    <p>To adjust to zero absorbance</p> Signup and view all the answers

    What is the wavelength at which the absorbance is measured in the spectrophotometer?

    <p>550 nm</p> Signup and view all the answers

    What is the concentration of the albumin solution in Tube 1?

    <p>0 mg/ml</p> Signup and view all the answers

    How should the results of the plasma solution be expressed?

    <p>mg protein per ml of undiluted plasma</p> Signup and view all the answers

    What is the primary purpose of estimating protein content in plasma derived from blood in this practical?

    <p>To compare with the standard protein albumin</p> Signup and view all the answers

    What is the minimum number of peptide bonds required for the Biuret reaction?

    <p>Two</p> Signup and view all the answers

    Why is accurate protein concentration determination important in biochemical molecular research?

    <p>For all aspects of biochemical molecular research</p> Signup and view all the answers

    What is the significance of peptide bonds in protein determination?

    <p>They are used to estimate protein concentration</p> Signup and view all the answers

    What is the importance of completing the test on Blackboard?

    <p>To help the student prepare for other practicals in the module</p> Signup and view all the answers

    What is the purpose of using 5 mg of albumin as a 'standard' protein?

    <p>To compare with other proteins</p> Signup and view all the answers

    Why is protein estimation important in biological samples?

    <p>For determining the amount of enzyme in a solution</p> Signup and view all the answers

    How many principal methods of protein determination are there?

    <p>Five</p> Signup and view all the answers

    What is the focus of this practical?

    <p>Comparing the relative number of peptide bonds in casein and gelatin</p> Signup and view all the answers

    Why is this practical not assessed?

    <p>Because it is a preparation for other practicals in the module</p> Signup and view all the answers

    What is the molarity of the sodium hydroxide solution added to the biuret reagent?

    <p>1M</p> Signup and view all the answers

    What is the purpose of vortexing the mixture in each tube?

    <p>To create a homogeneous mixture for spectrophotometric analysis</p> Signup and view all the answers

    What is the unit of measurement for the y-axis of the graph?

    <p>Absorbance units</p> Signup and view all the answers

    What is the primary advantage of using a calibration curve?

    <p>To determine the concentration of proteins in unknown samples</p> Signup and view all the answers

    What is the purpose of adding 4 ml of biuret reagent to each tube?

    <p>To initiate the biuret reaction</p> Signup and view all the answers

    What is the primary assumption underlying the estimation of protein concentration in plasma?

    <p>The protein concentration is directly proportional to the absorbance</p> Signup and view all the answers

    What is the purpose of plotting a graph in Excel?

    <p>To create a calibration curve</p> Signup and view all the answers

    What is the significance of the 20-minute waiting period before measuring the absorbance?

    <p>To allow the biuret reaction to reach equilibrium</p> Signup and view all the answers

    What is the purpose of using a disposable pipette tip for each separate solution?

    <p>To prevent cross-contamination between solutions</p> Signup and view all the answers

    What is the primary application of the biuret reaction in biochemical research?

    <p>To estimate protein concentration in biological samples</p> Signup and view all the answers

    What is the primary advantage of using the nitrogen content method for protein estimation?

    <p>It can cope with insoluble samples</p> Signup and view all the answers

    What is the main limitation of the Biuret reaction?

    <p>It is influenced by free amino acids</p> Signup and view all the answers

    Why is the Lowry method more sensitive than the Biuret reaction?

    <p>It is enhanced by Folin Ciocalteau's reagent</p> Signup and view all the answers

    What is the purpose of using a heavy metal in protein estimation?

    <p>To separate proteins from non-protein nitrogenous substances</p> Signup and view all the answers

    What is the primary limitation of ultraviolet absorption method for protein estimation?

    <p>It is influenced by the amino acid composition of proteins</p> Signup and view all the answers

    What is the purpose of wearing gloves and safety spectacles in the laboratory?

    <p>To protect the skin and eyes from chemicals</p> Signup and view all the answers

    What is the primary advantage of the dye-binding method?

    <p>It is not influenced by free amino acids</p> Signup and view all the answers

    What is the primary purpose of using a fume cupboard in the laboratory?

    <p>To contain hazardous vapours</p> Signup and view all the answers

    What is the primary limitation of methods that depend on the reaction of individual amino acids?

    <p>They are dependent on the amino acid composition of proteins</p> Signup and view all the answers

    What is the purpose of containing liquid spills with a towel roll in the laboratory?

    <p>To contain hazardous chemicals</p> Signup and view all the answers

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