Times New Ro... 12A A Aa ?! St PROBLEM SET #1-Do everything! Due 2/19/18 You wan

Question

Times New Ro... 12A A Aa ?! St PROBLEM SET #1-Do everything! Due 2/19/18 You want to grow cells in medium enriched with a-tocopherol (a-TC) to determine if this antioxidant protects the cells from drug-induced damage. The solubility of a-TC in aqueous medium is negligible so you will be making a stock solution of 1 M ?-TC in 100% ethanol. Describe how you would make 10 ml of this stock solution (molecular weight of a-TC is 472.8). Describe how you would prepare a minimum of 10 ml each of ?-TC solutions containing 10 mM, 1 mM, 0.1 mM, and 0.01 mM. 1. 2. Describe how you would prepare 1 L of a buffer containing 10 mM imidazole (mw 68.06), 10 mM EGTA (mw 380.4), and 100 mM KCl (mw 74.56) 3. You have 10 mL of cell suspension containing 500,000 cellsmle You want to plate a 24 well plate with equal numbers of wells at 3 densities: 100,000 cells/well, 50,000 cells/well, and 10,000 cells/well at a volume of 1 ml per well. Briefly describe how you would do so. f1 172 words English (US) + 10

Explanation / Answer

1- Given that the molecular weight of the alpha-tocopherol is 472.8

We need to make 10 ml of 1M stock solution

So we will be needing X gm of alpha-tocopherol

Molarity M = Mass/Mw * 1000/volume in ml

1 = X/472.8 *1000/10

X= 4.728 gm

To prepare 10mM solution we will take 0.1ml of the 1M solution of alpha-tocopherol and dissolve it in 9.9 ml ethanol.

To prepare 1mM solution we will take 1ml of the 10mM solution of alpha-tocopherol and dissolve it in 9ml ethanol.

To prepare 0.1mM solution we will take 1ml of the 1mM solution of alpha-tocopherol and dissolve it in 9ml ethanol.

To prepare 0.01mM solution we will take 1ml of the 0.1mM solution of alpha-tocopherol and dissolve it in 9ml ethanol.

2- 10mM IMIDAZOLE -Its molecular weight is 68.06

Molarity M = Mass/Mw * 1000/volume in ml

10*10-3 = X/68.06 *1

680 milligram

10mM EGTA Molecular weight 380.4

10*10-3 = X/380.4 *1

3.8gms

100mM KCl Mw - 74.56

100*10-3 = X/74.56

7.456gm

3-

For 100000cells/well

We will dilute the cell solution in 1:5 ratio.

1ml cells from stock solution i.e. 500000cell/ml and add it in 4 ml of DMEM.

This will give us 100000 cells/ml.

for 50000cells/ml

We will dilute the cell solution in 1:10 ratio

1ml cells from stock solution i.e. 500000cell/ml and add it in 9 ml of DMEM.

This will give us 50000 cells/ml.

For 10000cells per ml

We will dilute the cell solution in 1:50 ratio.

100 microlitre cells from stock solution i.e. 500000cell/ml and add it in 4.9 ml of DMEM.

This will give us 10000 cells/ml.

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