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Category: Science and Technology
Date Submitted: 09/19/2011 09:01 AM
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Title : Experiment 2- Estimation of protein concentration
Objectives :
1. To prepare a standard curve of absorbance versus protein concentration by using Bovine Serum Albumin (BSA).
2. To determine BSA concentration in two sample solutions.
3. To determine protein concentration in apple juice.
Materials :
Bradford Reagent, bovine serum albumin, apple juice sample, test tubes, cuvette, micropipette, tips.
Procedure :
As stated in Biological Science Practical Manual from page 11 to page 12.
Results :
Part 1: Preparation of standard curve of BSA
Table 3.1 Absorbance of BSA with different concentration
Tube | BSA 10µg/ml (µl) |
| Distilled water (µl) | Bradford reagent (µg/ml) | Concentration of BSA (µg/ml) | Absorbance at 595nm |
1 | 0 | 2000 | 500 | 0.00 | 0.000 |
2 | 100 | 1900 | 500 | 0.40 | 0.022 |
3 | 200 | 1800 | 500 | 0.80 | 0.037 |
4 | 300 | 1700 | 500 | 1.20 | 0.067 |
5 | 400 | 1600 | 500 | 1.60 | 0.089 |
6 | 500 | 1500 | 500 | 2.00 | 0.116 |
7 | 700 | 1300 | 500 | 2.80 | 0.183 |
8 | 900 | 1100 | 500 | 3.60 | 0.197 |
By using the formula, M1V1 = M2V2,
M1= Initial concentration of BSA
V1 = Volume of BSA
M2= Concentration of diluted BSA
V2 = Volume of diluted BSA
Tube 1 Tube 5
(10)(0) = M2 (2500) (10)(0) = M2 (2500)
M2 = 0 g/ml M2 = 1.60 g/ml
Tube 2 Tube 6
(10)(100) = M2 (2500) (10)(100) = M2 (2500)
M2 = 0.40 g/ml M2 = 2.00 g/ml
Tube 3 Tube 7
(10)(200) = M2 (2500) (10)(200) = M2 (2500)
M2= 0.80 g/ml M2= 2.80 g/ml
Tube 4 Tube 8
(10)(300) = M2 (2500) (10)(300) = M2 (2500)
M2 = 1.20 g/ml M2= 3.60 g/ml
Part 2: Determination of BSA concentration in two sample solutions &
Part 3: Determination of protein concentration of an apple juice
Table 3.2 Absorbance of sample solution A, B and C
Sample | Sample solution (µl) | Distilled water (µl) |...