Influence of different chemicals on the accuracy of the protein quantification assays

• Main Author: Tan, Hui Ying.
• Other Authors: School of Materials Science and Engineering
• Format: Final Year Project
• Language: English
• Published: 2012
• Subjects: DRNTU::Engineering::Materials::Biomaterials
• Online Access: http://hdl.handle.net/10356/50313
• Institution: Nanyang Technological University

Protein quantification assay is a fast and convenient method which is used to quantify extracted protein. Processes prior to extraction of protein involve chemicals which may affect the accuracy of the results. However these influences on the accuracy of the amount of protein quantified are not very well researched. Therefore, in this project, the chemicals involved in the prior processes such as Sodium Sulfide (Na2S), 2-Mercaptoethanol (2-ME), Urea and Tris(2-carboxyethyl)phosphine (TCEP) are experimented with the 2 different Protein Quantification Assays, Pierce 660nm (P660nm) and Bicinchoninic Acid (BCA), to find out the maximum amount or concentration of the chemicals that one can use in the processes. Bovine Serum Albumin is used as the standard for this project. From the results, the chemicals did show influence on the protein quantification and the maximum concentrations of the individual chemicals used for the respective assays are concluded. Furthermore, the possible effect of the influence is also discussed. In addition, both assays results were compared with the manufacturer’s protocol and P660nm assay showed a higher consistency than the BCA, hence P660nm might serve to be a more suitable assay than the latter.