You have a 25 ml enzyme sample that has a total of 100 micro-moles/min of activity...
You have a 25 ml enzyme sample that has a total of 100 micro-moles/min of activity
If you were to assay the enzyme, how much of it would you need in your assay in order to get an absorbance of 0.50 in a 30-minute assay.
The product E=13,800M-1cm-1 and you are using a standard 1mL cuvette
Beer-lambert law:
A=CIE
I=1
Homework Answers
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You have a 25 ml enzyme sample that has a total of 100
micro-moles/min of activity...
You have a 25 ml enzyme sample that has a total of 100 micro-moles/min of activity If you were to...
You have a 25 ml enzyme sample that has a total of 100 micro-moles/min of activity If you were to assay the enzyme, how much of it would you need in your assay in order to get an absorbance of 0.50 in a 30-minute assay. The product E=13,800M-1cm-1 and you are using a standard 1mL cuvette Beer-lambert law: A=CIE I=1
Question 3 10 points Save Ans It is important that you understand how to use the data generated b...
Question 3 10 points Save Ans It is important that you understand how to use the data generated by an absorbance based enzyme assay and relate this back to the enzyme activity Using the steps provided below. calculate the actvity of GPOH in the original preparation expressed as units ml tt two decimal places) under the following conditions: 75 uL glucose 6-phosphate dehydrogenase (G6PDH) was assayed in a reaction mixture with a total volume of 1.20 mL. The reaction was...
Sample Problem 1 TABLE 3-5 A Purification Table for a Hypothetical Enzyme Fraction volume (ml) Total...
Sample Problem 1 TABLE 3-5 A Purification Table for a Hypothetical Enzyme Fraction volume (ml) Total protein (mg) Activity (units) Procedure or step Specific activity (units/mg) 1,400 1. Crude cellular extract 2. Precipitation with ammonium sulfate 3. Ion-exchange chromatography 4. Size-exclusion chromatography 5. Affinity chromatography 10,000 3,000 400 10 32 100,000 96,000 80,000 60,000 45,000 200 600 15,000 • If affinity chromatography is the final step of purification, what is the yield and fold purification. Protein X has an absorptivity...
Q1: Say you have a patient serum sample. You follow the Lab 8 kinetic enzyme assay...
Q1: Say you have a patient serum sample. You follow the Lab 8 kinetic enzyme assay procedure for lactate dehydrogenase (LD) and obtain a slope of 0.0212 min^-1 at 340 nm. What is the patient's LD activity? Please show a work. Q2: If you fall down a flight of stairs, your serum CK activity will rise for the next two days or so. Explain why the serum CK activity increases. Activity (U/L) = (12650)(rate) One U/L activity is defined as...
Hello, I have been struggling with one of my biology maths questions. It involves calculating enzyme...
Hello, I have been struggling with one of my biology maths questions. It involves calculating enzyme activity and makes use of Beer's Law. Question: Calculate enzyme activity using your higher Vmax value. Molar extinction coefficient: 18,300 M-1 cm-1. Work out the concentrations of product/min and find the maximum rate in μmol/min. The equation I was given for the calculation: A=Ɛcd Vmax: 0.4 Abs/min Total sample volume: 3ml and enzyme was kept constant at 0.2 ml. Length of light path: 1cm...
(a) After spending a few weeks isolating the enzyme, you have enough pure enzyme to do...
(a) After spending a few weeks isolating the enzyme, you have enough pure enzyme to do some kinetic experiments, using a chromogenic substrate. The product produced by the enzyme from this substrate has an extinction coefficient of 20,500cm−1M−1 at 486nm, whereas the substrate does not absorb light significantly at this wavelength. For these experiments, you have used an enzyme concentration of 1.75µg/mL and monitored the change of absorbance at 486nm, using cuvettes with a path length of 1cm. The initial...
Does anyone know how to do this? Thanks in advance! Enzyme Activity (ml) Effect of pH...
Does anyone know how to do this? Thanks in advance!
Enzyme Activity (ml) Effect of pH on (amylase) enzyme activity. For determination of optimum assay pH of the enzyme reaction, 0.05 M Na2HPO4-NaH2PO4 buffer was used. The reaction was carried out for 10 min at 50°C in a shaking water. The enzyme activity was measured and the results are presented on graph, Bars represent means standard deviations for three replicates. 1. What do the error bars in this graph represent?...
i) When dihydrofolate reductase (10 Hg) was assayed in a total volume of 1 ml with 100 uM of dihydrofolate and 80 HM of deoxytubulosine, the enzyme activity was completely inhibited. After dialysi...
i) When dihydrofolate reductase (10 Hg) was assayed in a total volume of 1 ml with 100 uM of dihydrofolate and 80 HM of deoxytubulosine, the enzyme activity was completely inhibited. After dialysis of the assay mixture, enzyme activity was fully restored. What does this tell you about the mode of inhibition by deoxytubulosine irreversible or reversible? For your answer, illustrate how this type of inhibition could occur. [2 marks] v) Suggest TWO types of binding interactions that may take...
help please? this was the only other information given REPORT SHEET Determination of the Solubility-Product Constant...
help please?
this was the only other information given
REPORT SHEET Determination of the Solubility-Product Constant for a Sparingly Soluble Salt EXPERIMENT 8 A. Preparation of a Calibration Curve Initial (Cro121 0.0024 M Absorbance 5 mL Volume of 0.0024 M K Cro Total volume 1. I mL 100 mL 2. 100ML 3. 10 mL 100ml 4. 15 mL 100 ML Molar extinction coefficient for [CrO2) [Cro,2) 2.4x100M 12x1044 2.4810M 3.6810M 0.04) 2037.37 0.85 1.13 2. 3. Average molar extinction coefficient...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of this document. You will submit the worksheet via dropbox on Canvas by you assigned lab time the week of March 30th through April 3rd. A document with sample calculations of different concentrations is provided to you on canvas. Enzyme Kinetics Lab Buffer volume (mL) Enzyme Volume (ml) Substrate Volume (ml) TOTAL VOLUME (mL) 0.04 0.5 1.5 0.04 0.25 1.5 0.04 0.1 1.5 0.04 0.05...
Question 3 10 points Save Ans It is important that you understand how to use the data generated by an absorbance based enzyme assay and relate this back to the enzyme activity Using the steps provided below. calculate the actvity of GPOH in the original preparation expressed as units ml tt two decimal places) under the following conditions: 75 uL glucose 6-phosphate dehydrogenase (G6PDH) was assayed in a reaction mixture with a total volume of 1.20 mL. The reaction was...
Sample Problem 1 TABLE 3-5 A Purification Table for a Hypothetical Enzyme Fraction volume (ml) Total protein (mg) Activity (units) Procedure or step Specific activity (units/mg) 1,400 1. Crude cellular extract 2. Precipitation with ammonium sulfate 3. Ion-exchange chromatography 4. Size-exclusion chromatography 5. Affinity chromatography 10,000 3,000 400 10 32 100,000 96,000 80,000 60,000 45,000 200 600 15,000 • If affinity chromatography is the final step of purification, what is the yield and fold purification. Protein X has an absorptivity...
Does anyone know how to do this? Thanks in advance!
Enzyme Activity (ml) Effect of pH on (amylase) enzyme activity. For determination of optimum assay pH of the enzyme reaction, 0.05 M Na2HPO4-NaH2PO4 buffer was used. The reaction was carried out for 10 min at 50°C in a shaking water. The enzyme activity was measured and the results are presented on graph, Bars represent means standard deviations for three replicates. 1. What do the error bars in this graph represent?...
i) When dihydrofolate reductase (10 Hg) was assayed in a total volume of 1 ml with 100 uM of dihydrofolate and 80 HM of deoxytubulosine, the enzyme activity was completely inhibited. After dialysis of the assay mixture, enzyme activity was fully restored. What does this tell you about the mode of inhibition by deoxytubulosine irreversible or reversible? For your answer, illustrate how this type of inhibition could occur. [2 marks] v) Suggest TWO types of binding interactions that may take...
help please?
this was the only other information given
REPORT SHEET Determination of the Solubility-Product Constant for a Sparingly Soluble Salt EXPERIMENT 8 A. Preparation of a Calibration Curve Initial (Cro121 0.0024 M Absorbance 5 mL Volume of 0.0024 M K Cro Total volume 1. I mL 100 mL 2. 100ML 3. 10 mL 100ml 4. 15 mL 100 ML Molar extinction coefficient for [CrO2) [Cro,2) 2.4x100M 12x1044 2.4810M 3.6810M 0.04) 2037.37 0.85 1.13 2. 3. Average molar extinction coefficient...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of this document. You will submit the worksheet via dropbox on Canvas by you assigned lab time the week of March 30th through April 3rd. A document with sample calculations of different concentrations is provided to you on canvas. Enzyme Kinetics Lab Buffer volume (mL) Enzyme Volume (ml) Substrate Volume (ml) TOTAL VOLUME (mL) 0.04 0.5 1.5 0.04 0.25 1.5 0.04 0.1 1.5 0.04 0.05...
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