Pre-lab questions I (always write the answers to these in your lab notebook) soo

Question

Pre-lab questions I (always write the answers to these in your lab notebook) sooul ays write the answers to these in your lab notebook)/ co uF tisve tissue 1. How many microliters of homogenate will you work with for the experiment? How many 2. Before you load the SDS gel you will heat each homogenate sample. What is the effet of high 3. Ifyou were to make a l /5 dilution of 30 1 of your homogenate, how much water would you add milligrams of bovine tissue does that represent? SHOW YOUR CALCULATIONS. temperature on the structure proteins? to it? The table below lists a number of different possible consequences of a mutation in a cow's genome. Predict the kind of change you would expect to see in total protein concentration for each of the conditions listed using one of the following answers: "Higher for greater proteirn concentration in altered versus normal cells, "lower" if there would be lower protein concentration in altered versus normal cells, or "no detectable difference." THINK ABOUT THIS BEFORE YOU ANSWER. A low abundance A high abundance A single amino acid enzyme is missing structural protein is change prevents the A shortened nonfunctional missing breakdown of an enzyme is produced abundant structural protein

Explanation / Answer

1. It is given that the homogenate volume is 300 microliters. So as to find the amount of tissue present in the homogenate, you need to find out the concentration of the stock solution of BSA used. For example, if your BSA stock solution is 10% i.e (100mg/mL), then -

1 mL = 100 mg; so, 1 microliter = 100 microgram

Therefore, 300 microliters contain 30,000 micrograms or 30 mg of tissue.

3. Dilution (1/5) for 30 microliters homogenate will be carried out by taking 30 microliters of the homogenate and adding it to 120 microliters water to give 30/150 = 1/5 dilution (the dilution refers to the homogentate volume/ the total volume). Another way of doing the same is by taking 3 microliters of the homogenate and adding it to 12 microliters of water.

4. Total protein concentration will be affected as follows -

A low abundance enzyme is missing A high abundance structural protein is missing A single amino acid prevents the breakdown of an abundant structural protein A shortened, nonfunctional enzyme is produced no detectable change (since the enzyme was already in low abundance, major concentration changes will not occur). Lower (A highly abundant and hence important protein missiong will lead to lower concentrations). Lower (If the natural breakdown process does not occur, the protein concentration will be lower). no detectable change (since the enzyme produced is nonfunctional).

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