I am having issues trying to figure out how to find the protein concentration of

Question

I am having issues trying to figure out how to find the protein concentration of an unknown in a Bradford assay experiment.

I have no idea how to solve this problem. My professor just said "You take the x values determined from y = mx + b and they are the concentrations per tube that originally used for absorbances. Then you scale the x values up based on the original dilution factors you used to obtain those tubes in the first place, i.e. 10, 100 or 1000 dilution. For the y = mx + b, please input the absorbance as y, and solve for x from this." I am still lost.

How do I solve for this? How do I scale? I understand absorbance is y, but I don't know what m or b is in this equation and I don't know how data from the first set relates to the second set. I don't want the direct answer (why I didn't post my data, but I could come up with dummy data I guess if it helps solve the problem), but I'd like to know how to solve it. I have the absorbance factor for both the standard curve and the experimental, but not the protein for the experimental.

Explanation / Answer

While drawing a standard curve in this reaction, the concentration of the X axis, and mark the corresponding absorbance on Y xis.

Now join all the points which will give you a straight line.

Now when you will plot the graph in the excel sheet, you will get the line equation as y = mx+b

where m is the slope of the line and b is the y-intercept. And the y-intercept of this line is the value of y at the point where the line crosses the y axis.

So, slope is fixed for any graph.

And the Value of unknown concentration is placed in place of Y, Slope is known, constant b is known, So x can be calculated easily.

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