Publication Date

Spring 2011

Document Type

Project Summary

Degree Name

Master of Science

Department

Analytical Chemistry

First Advisor

Walter Henne, Jr., Ph.D.

Second Advisor

Patty Fu-Giles, Ph.D.

Third Advisor

Stephen Kent, M.B.A.

Abstract

Analysis of protein depends particularly on protein concentration. Protein concentration measurement is the most important part in the research work to conduct protein-related studies. Although there are many methods available for this purpose, each method has certain limitations. The aim of the experiment is to develop either new or modified analytical method for the analysis and detection of protein using newly introduced micro-plate reader equipped with Take-3 microplate with the help of Gen-5 software. An ideal assay should be simple and easy to carry out. Another aspect to be taken into consideration are low inference, stability of measured components and low protein to protein variation. This research work involves to measure protein concentration using various assays such as, absorbance at 280 nm, Bradford assay and BCA assay. Bradford and BCA assays are the most popular tools to quantify the protein sample. Bradford assay involves the measurement of absorbance at 595 nm. BCA assay involves measurement of absorbance at 562 nm. BSA was used as protein standard because it is highly pure and inexpensive. The overall strategy is to develop a robust assay that uses the least amount of sample and test reagents . Successful completion of this work will aid protein researchers in quick identication and analysis of proteins.