Hello Arooje Nasir

You will have to use the formula Y=mX+C.

This formula can be generated in excel. You need to plot your standard protein concentrations (BSA Std) on the X-axis and the corresponding absorbance values on the Y-axis using excel. In excel you need to choose the scatter plot. You need to add a trend line by right clicking on one of the points in the graph. This will result in the display of the equation Y=mX+C.

The excel will give you a linear line and generate the m and c values present in the equation. So, you can now calculate the X value (concentration) of your unknown sample by substituting the Y value which is the absorbance at 562 nm that you have already obtained for your unknown sample.

Therefore, the protein concentration of the unknown sample will be X = Y - C /m.

If you have diluted your unknown sample, then you will have to multiply X by the dilution factor to give you the protein concentration of the original undiluted sample. For instance, if the dilution factor is 10, then the final protein concentration of the unknown sample in the undiluted form will be 10X.

Hope this will help you.

Good Luck.

Dear Arooje Nasir You have to use increasing concentration of known protein (BSA) to prepare a standard curve. After plotting in excel you will get straight line, click on this graph and tick " display equation on chart" and "display R square value on chart" options. this will generate an equation in the form of Y=mx+c, where 'x' is the protein concentration and 'y' is the absorbance value of your unknown samples. if you dilute your sample you have to multiple the dilution factor to the value of 'X.

Good luck

https://www.researchgate.net/post/How_to_calculate_sample_protein_based_on_standard_protein_curve_in_BCA

https://ww2.chemistry.gatech.edu/~lw26/course_Information/4581/techniques/bradford/bradford.html

https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0057-Read-std-curves.pdf

Thanks everyone for providing valuable information and guidance,