ERIC Number: EJ1033345
Record Type: Journal
Publication Date: 2014-Jul
Abstractor: As Provided
Reference Count: 31
Rapid and Adaptable Measurement of Protein Thermal Stability by Differential Scanning Fluorimetry: Updating a Common Biochemical Laboratory Experiment
Johnson, R. Jeremy; Savas, Christopher J.; Kartje, Zachary; Hoops, Geoffrey C.
Journal of Chemical Education, v91 n7 p1077-1080 Jul 2014
Measurement of protein denaturation and protein folding is a common laboratory technique used in undergraduate biochemistry laboratories. Differential scanning fluorimetry (DSF) provides a rapid, sensitive, and general method for measuring protein thermal stability in an undergraduate biochemistry laboratory. In this method, the thermal denaturation of multiple different proteins is determined in parallel using a reverse-transcription polymerase chain reaction (RT-PCR) machine and a hydrophobic dye that differentially binds to proteins in non-native conformations. The utility of this methodology is illustrated by the measurement of differential protein stability in microplate volumes, in triplicate, with small protein samples. These characteristics make DSF measurement of protein stability adaptable to use with noncommercial protein samples. The methodology is also expandable to quantitating protein stability under a wide variety of solution conditions. The rapid setup and analysis of DSF experiments not only provides advanced undergraduates with experience in a fundamental biochemical technique but also provides the adaptability for use in inquiry-based laboratories and in independent research projects.
Descriptors: Biochemistry, Science Experiments, Laboratory Experiments, Science Activities, Undergraduate Students, College Science, Science Instruction
Division of Chemical Education, Inc and ACS Publications Division of the American Chemical Society. 1155 Sixteenth Street NW, Washington, DC 20036. Tel: 800-227-5558; Tel: 202-872-4600; e-mail: firstname.lastname@example.org; Web site: http://pubs.acs.org/jchemeduc
Publication Type: Journal Articles; Reports - Descriptive
Education Level: Higher Education; Postsecondary Education
Authoring Institution: N/A