Dilute BSA Standards

1. Label 9 tubes 1-9
2. Combine the components of the table below to generate appropriate concentration of solutions

Solution (ml)	BSA Standard Dilution						Unknown		Blank
	1 0.1mg/ml	2 0.2mg/ml	3  0.4mg/ml	4 0.6mg/ml	5  0.8mg/ml	6 1.0mg/ml	7	8	9 0.0mg/ml
ml BSA	0.1	0.2	0.4	0.6	0.8	1	–	–	–
ml H2O or ml Buffer	0.9	0.8	0.6	0.4	0.2	–	–	–	1
ml Sample	–	–	–	–	–	–	1	1	–
ml Biuret	4	4	4	4	4	4	4	4	4

3. Place tube 6 (1mg/ml) into a cuvette and measure absorbance (A) in the SpectroVis Plus. This will find the peak absorbance value.
4. Enter the LabQuest 2 application and press on the green Start button to generate a full spectrum
   • tap on the file cabinet icon to store this data
5. On the Meter Screen, tap on Mode
   1. Change the mode to “Events with Entry”
   2. Enter the Name: Concentration
   3. Enter Units: mg/ml
   4. Select OK
   5. If message appears about saving run, choose Discard
6. Sequentially read each sample at the stored wavelength (between A_540nm-A_600nm) and record values in table below

Test Tube	BSA (mg/ml)	Absorbance 540 nm
1	0.1
2	0.2
3	0.4
4	0.6
5	0.8
6	1.0
7	—
8	—

7. Plot each BSA dilution in plot.ly as a scatterplot
8. Generate best-fit line for these standards with the equation of the line
9. Use the equation of the line to estimate the concentration of the unknown sample.

Scatterplot Tutorial

https://youtu.be/bxGFS-RukZY