Homework Answers
the standard curve is drawn to estimate the concentration of nitrophenol in nmole/ml based on absorbance values or we can also say that,
this standard curve of absorbance is use to estimate the concentration of nitrophenol based on the optiocal density, OD410 that is absorbance at 410 nm.
thus according to this, the correct option for first question is OPTION A, that is on standard curve nitrophenol concentration is reported on the x axis,
and the correct option to second question is OPTION D, that is on standard curve absorbance at 410nm is reported on the y axis.
in third question,
for determining initial velocity the amount of product i.e. nitrophenol formed is plotted as a function of time and concentration is determined from the slope in the beginning of the reaction that is when the amount of product is increasing in the linear manner.
hence for question third the correct answer is OPTION D.
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In the standard curve for nitrophenol generated for use in this experiment, what is reported on...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV Absorbance 405 nm, was obtained If the UV absorbance 405 nm within the 1.5 mL-cuvette was measured as 0.144 in one of the trials during the experiment, determine the amount of pNP (in mg) produced. Assume total volume of reaction mixture stays at 1.5 mL at all times. (Round up to two decimal places) Molecular Weight of PNP: 139.11 g/mol PNP Concentration UV Absorbancem...
Is the standard curve reliable? What range is most reliable? Absorbance at 410 0.2 8 100...
Is the standard curve reliable?
What range is most reliable?
Absorbance at 410 0.2 8 100 120 p-nitrophenol conc in sample (nmol/mg gart A Concentration "
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs...
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs samples of known BSA concentrations. Now you need to determine the amount of protein in two mitochondria samples taken from fish, one taken from fish under normal conditions and the other taken from fish under oxidative stress. The absorbance reading for the normal fish fraction was A595=175, and the absorbance reading for the stressed fish fraction was A595= 275 Bradford Assay Standard Curve y...
Spectrophotometry help!? Would measuring the absorbance of the standard solutions used to prepare a calibration curve...
Spectrophotometry help!? Would measuring the absorbance of the standard solutions used to prepare a calibration curve and the unknown sample using a wavelength 10 nm higher or lower than lambda max provide an accurate concentration value for the unknown? Explain. If in an experiment, the pathlength of the light is 1 cm, and the concentration unit of the standard solutions is reported as mM, what would the units of the molar extinction coefficient, ε, be?
please use these results of a growth curve experiment to answer the following questions Questions 1)...
please use these results of a growth curve experiment to
answer the following questions
Questions 1) Plot the OD600 readings and cell concentration for each time point on a scatterplot with a line connecting the points. Have the measured data (OD600 and cell concentration) on the y-axis against the time on the x-axis using Excel (or other computer based data/graphing program). It is helpful to set the y-axis on a log scale for the cell concentration measurements. 2) Can you...
1- What is a standard curve and how is it used in biochemistry? 2- Explain how...
1- What is a standard curve and how is it used in biochemistry? 2- Explain how the Lambert-Beer equation is applied in this experiment. 3- Why is a spectrophotometer used to obtain data? 4- Based on the experimental data provided, estimate the amount of amino acid in the given unknown solution by Ninhydrin method. SI No. Amount of amino acid [pg) OD at 570nm 1 2 3 4 5 6 Volume of standard amino acid solution (ml) Blank 0.2 0.4...
. ous enzyme 'happyase catalyzes t the conversion of SAD to HAPPY with a Keat 100...
. ous enzyme 'happyase catalyzes t the conversion of SAD to HAPPY with a Keat 100 sec 40 35 30 25 20 10 10 20 30 40 50 60 70 90 100 110 ISADI QIM A kinetic experiment was conducted and the intitial velocity of the reaction as a function of SAD concentration is shown above. The initial velocity at 1 mM [SAD] was measured to be 40 nM/sec. a) What is the total [happyase] used above (make sure to...
THE DETERMINATION OF IRON BY SPECTROPHOTOMETRY INTRODUCTION In this experiment, the red-orange colored complex formed between...
THE DETERMINATION OF IRON BY SPECTROPHOTOMETRY INTRODUCTION In this experiment, the red-orange colored complex formed between iron(II) and 1,10- phenanthroline (Eqn.) is used in determination of iron by spectrophotometry. Fe+3PhenH Fe(Phen)2 +3H red-orange (A 512 nm) An excess of reducing reagent, such as hydroxylamine or hydroquinone, is often used to reduce and maintain iron in +2 oxidation state. The complex, once formed, is very stable, and can be stored for a long time. Required Reading: Skoog and West (9E): Chapter...
Beer’s Law Objective : We will explore an application of absorption spectroscopy using calibration curves and...
Beer’s Law Objective : We will explore an application of absorption spectroscopy using calibration curves and Beer’s Law. Use the “LAB : HOW TO…” link from the class website if you need help with how to use balance, Bunsen burner… and such. Introduction: You may write this information in your lab notebook for your own reference. It can’t be cut and pasted. Different solutions have different spectral properties. In this portion of the experiment those properties will be utilized to...
Experiment # Starting Substrate Concentration Actual Substrate Concentration Substrate Added (ml) Buffer Added (ml) 1.5 mm...
Experiment # Starting Substrate Concentration Actual Substrate Concentration Substrate Added (ml) Buffer Added (ml) 1.5 mm 1.5 1.0 mm Nm 1.0 0.5 mm 0.50 0.250 mm 0.250 0.125 mm 0.120 1.38 0.060 MM 0.060 1.44 0.030 mm 0.030 1.47 0.015 mm 0.015 1.485 *Actual substrate concentration is calculated by taking the starting substrate concentration into the final volume of each reaction after enzyme has been added. Note that the total volume for each reaction prior to the addition of enzyme...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV Absorbance 405 nm, was obtained If the UV absorbance 405 nm within the 1.5 mL-cuvette was measured as 0.144 in one of the trials during the experiment, determine the amount of pNP (in mg) produced. Assume total volume of reaction mixture stays at 1.5 mL at all times. (Round up to two decimal places) Molecular Weight of PNP: 139.11 g/mol PNP Concentration UV Absorbancem...
Is the standard curve reliable?
What range is most reliable?
Absorbance at 410 0.2 8 100 120 p-nitrophenol conc in sample (nmol/mg gart A Concentration "
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs samples of known BSA concentrations. Now you need to determine the amount of protein in two mitochondria samples taken from fish, one taken from fish under normal conditions and the other taken from fish under oxidative stress. The absorbance reading for the normal fish fraction was A595=175, and the absorbance reading for the stressed fish fraction was A595= 275 Bradford Assay Standard Curve y...
please use these results of a growth curve experiment to
answer the following questions
Questions 1) Plot the OD600 readings and cell concentration for each time point on a scatterplot with a line connecting the points. Have the measured data (OD600 and cell concentration) on the y-axis against the time on the x-axis using Excel (or other computer based data/graphing program). It is helpful to set the y-axis on a log scale for the cell concentration measurements. 2) Can you...
1- What is a standard curve and how is it used in biochemistry? 2- Explain how the Lambert-Beer equation is applied in this experiment. 3- Why is a spectrophotometer used to obtain data? 4- Based on the experimental data provided, estimate the amount of amino acid in the given unknown solution by Ninhydrin method. SI No. Amount of amino acid [pg) OD at 570nm 1 2 3 4 5 6 Volume of standard amino acid solution (ml) Blank 0.2 0.4...
. ous enzyme 'happyase catalyzes t the conversion of SAD to HAPPY with a Keat 100 sec 40 35 30 25 20 10 10 20 30 40 50 60 70 90 100 110 ISADI QIM A kinetic experiment was conducted and the intitial velocity of the reaction as a function of SAD concentration is shown above. The initial velocity at 1 mM [SAD] was measured to be 40 nM/sec. a) What is the total [happyase] used above (make sure to...
THE DETERMINATION OF IRON BY SPECTROPHOTOMETRY INTRODUCTION In this experiment, the red-orange colored complex formed between iron(II) and 1,10- phenanthroline (Eqn.) is used in determination of iron by spectrophotometry. Fe+3PhenH Fe(Phen)2 +3H red-orange (A 512 nm) An excess of reducing reagent, such as hydroxylamine or hydroquinone, is often used to reduce and maintain iron in +2 oxidation state. The complex, once formed, is very stable, and can be stored for a long time. Required Reading: Skoog and West (9E): Chapter...
Experiment # Starting Substrate Concentration Actual Substrate Concentration Substrate Added (ml) Buffer Added (ml) 1.5 mm 1.5 1.0 mm Nm 1.0 0.5 mm 0.50 0.250 mm 0.250 0.125 mm 0.120 1.38 0.060 MM 0.060 1.44 0.030 mm 0.030 1.47 0.015 mm 0.015 1.485 *Actual substrate concentration is calculated by taking the starting substrate concentration into the final volume of each reaction after enzyme has been added. Note that the total volume for each reaction prior to the addition of enzyme...
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