Bioscience Experiment 2 Protein Concentration
Experiment 2: Estimation of protein concentration
Name: Wong Jiun Hao
ID: 00000011136
Cohort: BM114
Module: Biological Science
Table of Contents
Title 1
Table of Contents 2
Introduction 3
Objectives 4
Procedure 4
Results 5-6
Discussion 7-8
Conclusion 9
References 9
Introduction
The concentration of protein in a sample can be obtained by using the Bradford protein assay method. The method requires a spectrophotometer and it is easily operated. The Bradford protein assay is a colorimetic assay based on the interaction of the blue dye “Coomassie” and the protein. The blue ye firsts donates free electron to the protein and causes the protein to lose its native state and exposing it hydrophobic pockets. These hydrophobic pockets then binds to the Coomassie dye through Van Der Wall forces. The binding protein then stabilizes the blue form of the Coomassie dye. Hence, the absorbance reading can be obtained and used to measure the protein concentration which is the amount of protein complex present in the solution. To find the protein concentration from the standard curve. First, draw the line of best fit and include the equation into the graph. The R2 will be as close to 1 as possible. Apply the Beer Lambert Law A= λbc in which A is the measured absorbance, λ is the slope of the standard curve, b is the length of the cuvette, and c is the concentration being determined.
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.
Procedure
Part1
7 test tubes were prepared and labeled. The appropriate amount of BSA and distilled water were pipetted into the respective test tubes according Table 3.1. 500µl of Branford Reagent were added into all the test tubes. The test tubes were all vortexed and mixed.
Name: Wong Jiun Hao
ID: 00000011136
Cohort: BM114
Module: Biological Science
Table of Contents
Title 1
Table of Contents 2
Introduction 3
Objectives 4
Procedure 4
Results 5-6
Discussion 7-8
Conclusion 9
References 9
Introduction
The concentration of protein in a sample can be obtained by using the Bradford protein assay method. The method requires a spectrophotometer and it is easily operated. The Bradford protein assay is a colorimetic assay based on the interaction of the blue dye “Coomassie” and the protein. The blue ye firsts donates free electron to the protein and causes the protein to lose its native state and exposing it hydrophobic pockets. These hydrophobic pockets then binds to the Coomassie dye through Van Der Wall forces. The binding protein then stabilizes the blue form of the Coomassie dye. Hence, the absorbance reading can be obtained and used to measure the protein concentration which is the amount of protein complex present in the solution. To find the protein concentration from the standard curve. First, draw the line of best fit and include the equation into the graph. The R2 will be as close to 1 as possible. Apply the Beer Lambert Law A= λbc in which A is the measured absorbance, λ is the slope of the standard curve, b is the length of the cuvette, and c is the concentration being determined.
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.
Procedure
Part1
7 test tubes were prepared and labeled. The appropriate amount of BSA and distilled water were pipetted into the respective test tubes according Table 3.1. 500µl of Branford Reagent were added into all the test tubes. The test tubes were all vortexed and mixed.
References: 1. http://www.sigmaaldrich.com/life-science/proteomics/protein-quantitation/bradford-reagent.html 2. Bradford protein assay. (2014, October 28). In Wikipedia, The Free Encyclopedia. Retrieved 01:29, November 17, 2014, from http://en.wikipedia.org/w/index.php?title=Bradford_protein_assay&oldid=631530871