Title

Accelerated Western Blotting Lab for Biochemistry I Laboratory Curriculum

Abstract

The purpose of this current study was to design a Western Blotting procedure for the Biochemistry Laboratory curriculum at NMU. The methodology consists of gel electrophoresis, transfer of proteins to a membrane surface, and then a series of immunodetection steps that utilize antibodies to recognize and bind to a specific antigen protein. The final steps involve an enzyme, which, when reacted with substrate, allows for the detection of a band on the Western Blot. These bands act as flags, singling out the desired protein. Western Blotting is an important skill for students to learn and is not currently part of the Biochemistry I Laboratory curriculum. Traditionally, this process takes approximately three days to complete, which makes it very difficult to incorporate into the laboratory curriculum schedule. Recently, BioRad has created a piece of equipment, the Turbo Transfer system, that can perform a Western transfer in just seven minutes, which reduces the needed lab time by one day. Using this new piece of technology, it was possible to design a Western Blot experiment for students. To achieve this overall goal, the following objectives were identified: verify the antibody chosen could identify the protein of interest, compare the traditional transfer protocol to the faster transfer system, and optimize the Western Blot protocol for use in the teaching lab environment. All objectives have been met and study results have been proven to be reproducible through repeated testing. This project will conclude with the completion of a performable Western Blotting lab procedure for students.

Class Standing

Freshman

Department

Chemistry

Faculty Advisor

Crystal Keso

Faculty Advisor Email

ckeso@nmu.edu

Date

2020

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