Objective

 how to prepare a tissue protein extract and use a protein assay reagent to measure the protein
concentration.
 Learn the theory and practice of absorbance spectrophotometry.
 how to prepare and use a standard curve and absorption spectrum.

Materials

 Stock solution of Bovive Serum Albumin (BSA): 10mg/ml

 Deionized water (dH2O)
 Test tube
 Test tube track
 Pipette
 Biuret reagent
 Spectrophotometer

Methodology

1. 9 test tube prepared with this item.

Test tube Gelatin, H2O, mL 2 mL of 15 minutes

mL Biuret incubation
1 0.0 1.0
2 0.1 0.9
3 0.2 0.8
4 0.3 0.7
5 0.4 0.6
6 0.5 0.5
7 0.6 0.4
8 (Kitkat) 1.0 5.0
9(Cadbury) 1.0 5.0

2. Incubate the test tube 15 minutes at room temperature.

3. The test tube entered in Spectrophotometer 580nm.
4. The absorbance recorded.
Results

Protein Absorbance
concentration,
(mg/mL)
0 0.000
1 0.180
2 0.277
3 0.391
4 0.526
5 0.619
6 0.684
x2 0.627
x1 0.907