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Protein Purification 2: Size Exclusion Chromatography

This resource addresses the following Core Concepts of Vision and Change: 1. EVOLUTION 2. STRUCTURE AND FUNCTION 4. PATHWAYS AND TRANSFORMATIONS OF ENERGY AND MATTER This resource can help students develop the following Core Competencies of Vision and Change: 1. ABILITY TO APPLY THE PROCESS OF SCIENCE 2. ABILITY TO USE QUANTITATIVE REASONING 5. ABILITY TO COMMUNICATE AND COLLABORATE WITH OTHER DISCIPLINES These materials are intended to be part of a Vision and Change Collection about enzyme purification. This lab activity continues the purification of adenosine deaminase (ADA). The salt pellet is loaded onto a size exclusion column. This column serves as a desalting step by removing the ammonium sulfate and as a purification step by fractionating the proteins based on size. Enzyme activity is located using a QUALITATIVE spectrophotometric assay, and active fractions are pooled for dialysis. Buffers for the ion exchange chromatography step are chosen, prepared, and used for dialysis. Students work in teams (typically of 3-4) to complete the objectives and analyze the data. The pre-lab lecture component uses SCALE-UP to engage the students and prepare them for lab. Students watch “lecture chunks” (created by breaking up my traditional 60 minute Ppt lectures into 3-5 minute video clips) before the pre-lab meeting: these short presentations about biochemical theory and equipment set-up allow the instructor to “flip the classroom” (i.e., learn before lecture) so class time can incorporate more student-centered activities. I have additional Ppt slides with clicker questions that I use during the class discussion before lab that complement the student-centered activities. These are tested materials currently used in a seven day advanced undergraduate laboratory course (BIOC 311: Advanced Experimental Biosciences) that is targeted at juniors and seniors.

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Resource Type: Digital Presentation (Powerpoint), Short Movies, Assessment: other, Laboratory or Hands-On Activity, Laboratory Manual, Lecture / Lecture Outline / Lecture Notes, Teaching Strategies & Guidelines, Tutorial
Audience Level: Undergraduate upper division (Grades 15-16), Graduate

Author and Copyright


Authors and Editors: PhD Beth Beason-Abmayr of Rice University Biochemistry and Cell Biology
Publisher: Rice University Biochemistry and Cell Biology
Copyright and other restrictions: No
Cost: No

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Collection:
American Physiological Society


     
   

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