Quantitative Detection of Protein (SpectroVis Plus)

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

0.4mg/ml

4

0.6mg/ml

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

  1. Place tube 6 (1mg/ml) into a cuvette and measure absorbance (A) in the SpectroVis Plus. This will find the peak absorbance value.
  2. 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
  3. 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
  4. Sequentially read each sample at the stored wavelength (between A540nm-A600nm) 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

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

Scatterplot Tutorial

https://youtu.be/bxGFS-RukZY