High-throughput differential scanning fluorimetry of GFP-tagged proteins
Other Publication ResearchOnline@JCUAbstract
Differential scanning fluorimetry is useful for a wide variety of applications including characterization of protein function, structure–activity relationships, drug screening, and optimization of buffer conditions for protein purification, enzyme activity, and crystallization. A limitation of classic differential scanning fluorimetry is its reliance on highly purified protein samples. This limitation is overcome through differential scanning fluorimetry of GFP-tagged proteins (DSF-GTP). DSF-GTP specifically measures the unfolding and aggregation of a target protein fused to GFP through its proximal perturbation effects on GFP fluorescence. As a result of this unique principle, DSF-GTP can specifically measure the thermal stability of a target protein in the presence of other proteins. Additionally, the GFP provides a unique in-assay quality control measure. Here, we describe the workflow, steps, and important considerations for executing a DSF-GTP experiment in a 96-well plate format.
Journal
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Publication Name
Targeting Enzymes for Pharmaceutical Development
Volume
2089
ISBN/ISSN
1064-3745
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Pages Count
17
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Publisher
Springer
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Publisher Location
New York, NY, USA
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DOI
10.1007/978-1-0716-0163-1_5