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ERIC Number: EJ1126366
Record Type: Journal
Publication Date: 2017
Pages: 9
Abstractor: As Provided
ISSN: ISSN-1470-8175
Using an FPLC to Promote Active Learning of the Principles of Protein Structure and Purification
Robinson, Rebekah L.; Neely, Amy E.; Mojadedi, Wais; Threatt, Katie N.; Davis, Nicole Y.; Weiland, Mitch H.
Biochemistry and Molecular Biology Education, v45 n1 p60-68 Jan-Feb 2017
The concepts of protein purification are often taught in undergraduate biology and biochemistry lectures and reinforced during laboratory exercises; however, very few reported activities allow students to directly gain experience using modern protein purification instruments, such as Fast Protein Liquid Chromatography (FPLC). This laboratory exercise uses size exclusion chromatography (SEC) and ion exchange (IEX) chromatography to separate a mixture of four different proteins. Students use an SEC chromatogram and corresponding SDS-PAGE gel to understand how protein conformations change under different conditions (i.e. native and non-native). Students explore strategies to separate co-eluting proteins by IEX chromatography. Using either cation or anion exchange, one protein is bound to the column while the other is collected in the flow-through. In this exercise, undergraduate students gain hands-on experience with experimental design, buffer and sample preparation, and implementation of instrumentation that is commonly used by experienced researchers while learning and applying the fundamental concepts of protein structure, protein purification, and SDS-PAGE.
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Publication Type: Journal Articles; Reports - Descriptive
Education Level: Higher Education; Postsecondary Education
Audience: N/A
Language: English
Sponsor: N/A
Authoring Institution: N/A
Grant or Contract Numbers: N/A