Describe how you would make the following solution containing two enzymes. 1 mL total volume in Buffer X with 10nM of enzyme A and 50nM of enzyme B You are given a solution of enzyme A at 1 mg/mL and a solution of enzyme B at 1 mg/mL. The molecular weight of enzyme A is 75,000 g/mol and the molecular weight of enzyme B is 130,000 g/mol.
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Describe how you would make the following solution containing two enzymes.
1 mL total volume in Buffer X with 10nM of enzyme A and 50nM of enzyme B
You are given a solution of enzyme A at 1 mg/mL and a solution of enzyme B at
1 mg/mL. The molecular weight of enzyme A is 75,000 g/mol and the molecular weight of enzyme B is 130,000 g/mol.
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- You have a stock solution containing 1.5 mM of substrate in borate buffer pH 9.5. You transfer 1.25 mL of this stock into a cuvette and then add 250 µl of enzyme solution to initiate the reaction. What will be the final concentration of substrate in the cuvette following addition of the enzyme? 1.5 mM 1.25 mM 0.0075 mM 7.5 mM 1250 mMYou need to make a protein buffer of: . • • 100 mM NaCl 25 mM Tris 8 5% w/v glycerol • 2 mM DTT Your stock solutions are: 5 M NaCl • 2 M Tris 8 • 70% w/v glycerol • DTT @ 154.25 g/mol How would you make a 1L protein buffer Solution? Show your work and describe.2.5 2 1.5 1 0.5 0 = Mike has determined that enzyme he is attempting to purify has an isoelectric point of 4.5 (pl: 4.5). He has decided to examine anion exchange chromatography as a potential purification step. He tested out using 2 different buffer and linear NaCl gradient on HPLC (as what you did in lab). His results are shown below. pH 8 data 1 3 4 5 7 9 10111315171920212325272930 Fraction number -A280 Enzyme activity 2.5 2 1.5 1 0.5 0 pH 5 data 1 3 4 5 7 9 10111315171920212325272930 Fraction number -A280 --Enzyme activity Which buffer should Mike uses for his purification (both buffer has pH higher than enzyme's pl)? Why
- Calculate the amount of rice bran (12.5% CP), corn (8.5% CP), soybean meal (49% CP) and copra meal (20% CP) in formulating 100 kg diet with 20% CP. Assume that the energy mixture will have a ratio of 2:1 and protein ingredients proportion is 1:3. Show your solution:A researcher is preparing a reaction mixture to test the activity of a protein. They combine the required reaction components, which contained in a final 100 ml reaction volume 200 mM NaCl, unknown concentrations of acetic acid and acetate anions and a total [H+] concentration of 790 ricromolar. Can you determine the pH of the solution? Provide the answer to one decimal place. Note: You may need to round the numbers to get the required answer. 100 Strips pH indicator strips non-bleeding pH pH 0-14 EM-Reagents Dip in-read while still moist. immerse in weakly-buffered solutions une there is no further color change (1-10 min) 3 7 5 6The isoelectric point of eIF4a is 5.02. Students are given a sample of cell lysate containing eIF4A at pH 7.4. They are also provided with two buffers of pH 7.4 to use for ion exchange chromatography. One buffer has a very high salt concentration (1 M), the other has a low salt concentration (0.1 M). Mariela decides to use an anion exchange column, while Ashok chooses a cation exchange column. Who made the better decision? Provide a detailed explanation of why one student will end up with a purer sample of eIF4A. Which buffer will you use while washing away impurities (high or low salt), and which would you use to remove eIF4A from the column? Explain your choice.
- You are supplied with the following: NaCl (Mr = 58.443 g/mol) 250 mM Tris-Cl, pH 8 solution (1 Litre) EDTA, sodium salt (Mr = 380.2 g/mol) 13 % Sodium dodecyl sulfate solution Proteinase K solution (150 mg dissolved in 1 ml ddH2O) You need a digestion buffer consisting of the following: 23.5 mM NaCl 750 uM Tris-Cl, pH 8 38 mM EDTA, pH 8 1.2 % Sodium dodecyl sulfate 1.75 mg/ml proteinase K Calculate: How will you prepare 750 ml of the digestion buffer? Show all your steps and calculations and remember to explain how exactly you will make it up.The enzyme urease is widely used for determining urea concentration in blood. The Michaelis constant for urease at room temperature is 2.0 mM and k2 = 2.8 × 104 s-1 at pH = 7.5. Based on the kinetic experiment, when 0.34 mM urease was used, the initial rate of the reaction was determined to be 3.49 M/sec. What is the concentration of urea in blood in unit of mM? Please keep your answer to two decimal place. Please convert all concentration unit to mM in calculation.Highlight your values of A,B,C and D. For your question: A mL of B mol/L sodium phosphate solution is combined with C mL of D mol/L calcium bicarbonate. (Calcium bicarbonate is soluble.) A mL B mol/L C mL D mol/L 75.0 ml 0.300 67.5 0.350 Before you begin your reaction, you must accurately produce 1.500 L of your sodium phosphate solution from sodium phosphate trihydrate solid. Write out a procedure to explain all the steps you will take in the lab when making the solution to ensure that your solution concentration is accurate. Please include calculations that show the required mass of solid. Also include the correct names of all equipment used.
- NaCl (Mr = 58.443 g/mol) 2.5 M Tris-Cl, pH 8 oplossing / solution (1 Litre) EDTA, sodium salt (Mr = 380.2 g/mol) 10 % Sodium dodecyl sulfate solution Proteinase K solution (50 mg dissolved in 1 ml ddH2O) You need a digestion buffer consisting of the following: 15 mM NaCl 75 mM Tris-Cl, pH 8 16 mM EDTA, pH 8 0.8 % Sodium dodecyl sulfate 0.75 mg/ml proteinase K Calculate: How will you prepare 500 ml of the digestion buffer? Show all your steps and calculations and remember to explain how exactly you will make it up.You have just used up the last of a 1.2 % m/m solution of agarose in water. You require another 100mls. How many grams of Agarose do you require? Note 1ml of water=1g What is the % w/v of a 100ml solution containing 5g of NaCl?1.) A sample contains 0.003 mg of NaOH. How many micrograms (μg) is this? 2.) An experiment was performed in which 400 mL of 0.2 M urea was used. Calculate the number of millimoles (mmol) of urea contained in this sample. 3.) How many micromoles (μmol) of ATP are present in 4 mL of 5 mM ATP solution..