Materials: Reaction Buffer 0.5 M Tris HCl pH 8.0 with 5 mM MgCl2 Alkaline phosphatase: 500...

Materials:

Reaction Buffer 0.5 M Tris HCl pH 8.0 with 5 mM MgCl2

Alkaline phosphatase: 500 μg/mL stock solution in the buffer above. Keep enzyme on ice!

p-nitrophenyl phosphate: 1000 μM and 10000 μM stock solutions in reaction buffer

1) Using the enzyme stock solution above and the dilution equation, calculate the volumes needed to prepare 2 mL of three enzyme dilutions consisting of 100 μg/mL, 125 μg/mL, and 150 μg/mL. Use the reaction buffer to make these dilutions.

2) Calculate the amount of substrate needed for each of the following concentrations 10 μM, 25 μM, 50 μM, 100 μM, 200 μM, 500 μM, 1000 μM, and 2000 μM using the two stock solution concentrations listed above. Use the 1000 μM stock solution to calculate dilutions from 10-200 μM and the 10,000 μM stock solution to calculate from 500 μM-2000 μM. The final volume is 1.0 mL (or 1000 μL) for each dilution. Also calculate the volume of buffer needed, given that we will add 100 μL of the enzyme solution and the final volume is 1.0 mL (or 1000 μL) for each dilution.

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queen_honey_blossom answered 2 years ago

1. Tris buffer a. 100 mM Tris-HCl, pH 8.0 b. 100 mM Tris-HCl, pH 6.0 Make...

1. Tris buffer a. 100 mM Tris-HCl, pH 8.0 b. 100 mM Tris-HCl, pH 6.0 Make this buffer by dissolving an appropriate amount (which you must calculate using the formula weight) of solid Tris in APPROXIMATELY 80 mL of water. Use the Henderson-Hasselbalch equation to calculate how much 1 M HCl is needed to achieve the final pH?

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Suppose you want to make 500 mL of a 0.20M Tris buffer at pH 8.0. On...

Suppose you want to make 500 mL of a 0.20M Tris buffer at pH 8.0. On the shelf in lab, you spot a bottle of 1.0M Tris at pH 6.8 and realize you can start with that to make his new buffer. Assuming you have 5.0M HCl and 5.0M NaOH at your disposal, how could you make this 0.20M Tris buffer at pH 8.0 from the 1.0M Tris, pH 6.8? (The pKa of Tris is 8.06.)

Given stock solutions of 25 mM nitrophenyl phosphate, and 1.5 M tris,pH 7.2, 7.5 mM MgCl2....

Given stock solutions of 25 mM nitrophenyl phosphate, and 1.5 M tris,pH 7.2, 7.5 mM MgCl2. If you mix 0.1ml of the nitrophenyl phosphate, 0.7 ml of the tris solution and 0.2 ml of water to bring the total volume to 1 ml, what is the final concentrations of each of the components in the final solution? Show work Nitrophenyl phosphate in mM Tris in M Magnesium chloride in mM

How would you make up 100 ml of 50 mM Tris-HCl buffer at pH 7.4, using...

How would you make up 100 ml of 50 mM Tris-HCl buffer at pH 7.4, using the 0.5 M Tris base stock solution, and any other required materials?

Calculate the pH of the 0.250 M NH3/0.5 M NH4Cl buffer system. What is the pH...

Calculate the pH of the 0.250 M NH3/0.5 M NH4Cl buffer system. What is the pH after the addition of 2.0 mL of 0.250 M NaOH to 18.0 mL of the buffer solution? After adding 10 more mL of 0.25 M NaOH what is the pH?

How you would prepare a 500 mL solution of 0.1M "tris" buffer at pH of 7.5...

How you would prepare a 500 mL solution of 0.1M "tris" buffer at pH of 7.5 starting with solid tris free base. The free base has a chemical forumla of (HOCH2)3-C-NH2 and a MW of 121.1 g/mol. The protonated amine form, tris-chloride, has a pKa of 8.10. To prepare the solution, 1.0M HCl will be used.

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Materials: Reaction Buffer 0.5 M Tris HCl pH 8.0 with 5 mM MgCl2 Alkaline phosphatase: 500...

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