Interpreting Protein standardized curve

[songl4a]

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[referee3]

Hello, just a quick question about protein concentration quantification.

I followed this procedure: https://www.piercenet.com/instructions/2160129.pdf
with the test tube procedure.

I used the chart on page 2 to dilute the BSA standard and then graphed the absorbance and got a polynomial equation. Then I took a mL of my unknown sample and followed the same exact chart for the dilutions and then found the absorbance of each dilution.

Was I supposed to only take 30 uL of each of my unknown samples and mix it with the dye instead of doing the serial dilution? Otherwise, I would only know the concentration of 1 sample right?

Furthermore, how do I convert the polynomial equation to solve in terms of X (concentration) so I can simply plug in the absorbance values (Y) of my unknown to find the concentration.

Thanks!

If you still need help with this pm me. I do these same assays all the time in lab with the same kit.