Please show how these answers were obtained. Thank you if you can help Justin ob

Question

Please show how these answers were obtained. Thank you if you can help

Justin obtains the following data for his BSA standard curve for a coomassie assay. He also tests the UK and then three dilutions: 1: 5, 1: 10 and 1: 50. The OD values for the UK and the three dilutions are respectively: 0.55, 0.45, 0.38, and 0.26. The equation for the trendline is y = 0.0275x + 0.117. a. Which dilution should Justin pick as the best measure of the concentration of the stock? 1: 50 b. How much protein (in mu g) is in the dilution? 5.2 mu g c. If Justin assayed 5 mu l of the dilution, what is the concentration? 1.04 mu g

Explanation / Answer

a) 1:50 is the third and last dilution. As the sample is diluting, the amount of original sample or concentration proteins in the stock become decrease , there by it is easy to analyse separately and get the exact accurate result. When the sample is highly concentrated without any dilution or with low dilution, the targeted product in the sample become cloudy and rush which will make the variations in the result.

b) For dilution 1:50, we got an OD value of 0.26.

From the values in the given table, we can understand the amount of protein corresponding to the OD values.

Accordingly,

OD value 0.23 = 4microgram

OD value 0.28 = 6 microgram

The difference between 0.23 and 0.28 is 0.05.

And difference between 4 and 6 is 2.

So for OD value 0.05 = 2 microgram

So for OD 0.01= 0.4

OD 0.03 = 1.2

We already know that for OD value 0.23 = 4microgram

Now we need to find 0.26 = how much.

Hence,

For OD value 0.23+0.03 = 4+1.2

That is , OD value 0.26 = 5.2 microgram of protein.

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