Rapid and Adaptable Measurement of Protein Thermal Stability by Differential Scanning Fluorimetry: Updating a Common Biochemical Laboratory Experiment
| Title: | Rapid and Adaptable Measurement of Protein Thermal Stability by Differential Scanning Fluorimetry: Updating a Common Biochemical Laboratory Experiment |
|---|---|
| Language: | English |
| Authors: | Johnson, R. Jeremy, Savas, Christopher J., Kartje, Zachary |
| Source: | Journal of Chemical Education. Jul 2014 91(7):1077-1080. |
| Availability: | 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: eic@jce.acs.org; Web site: http://pubs.acs.org/jchemeduc |
| Peer Reviewed: | Y |
| Page Count: | 4 |
| Publication Date: | 2014 |
| Document Type: | Journal Articles Reports - Descriptive |
| Education Level: | Higher Education Postsecondary Education |
| Descriptors: | Biochemistry, Science Experiments, Laboratory Experiments, Science Activities, Undergraduate Students, College Science, Science Instruction |
| DOI: | 10.1021/ed400783e |
| ISSN: | 0021-9584 |
| Abstract: | 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. |
| Abstractor: | As Provided |
| Number of References: | 31 |
| Entry Date: | 2014 |
| Accession Number: | EJ1033345 |
| Database: | ERIC |
| ISSN: | 0021-9584 |
|---|---|
| DOI: | 10.1021/ed400783e |
| Published in: | Journal of Chemical Education |
| Language: | English |