A High-Throughput Fluorescence Polarization-Based Assay for the SH2 Domain of STAT4
Abstract
:1. Introduction
2. Materials and Methods
2.1. Peptide Sequences
2.2. Cloning and Protein Expression
2.3. Fluorescence Polarization Assays
2.4. Calculation of Z’
3. Results and Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Peptide Sequence | Target Protein | IC50 [µM] or Inhibition at 40 µM [%] | Ki [µM] [a] |
---|---|---|---|
Ac-GpYLPQNID | STAT4 | 0.49 ± 0.04 µM | 0.22 ± 0.02 µM |
Ac-pYLPQTV-NH2 | STAT3 | 1.27 ± 0.10 µM | 0.60 ± 0.05 µM |
DTpYLVLDKWL | STAT5a/5b | 4.17 ± 0.13 µM | 1.99 ± 0.06 µM |
GpYDKPHVL | STAT1 | 31.4 ± 6.7 µM | 15.1 ± 3.2 µM |
GpYEEIP | Lck/Src | 37 ± 4 % inhibition | n/a |
MAGPMQSpTPLNGAKK | Plk1 PBD | 2 ± 2 % inhibition [b] | n/a |
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Berg, A.; Gräber, M.; Schmutzler, S.; Hoffmann, R.; Berg, T. A High-Throughput Fluorescence Polarization-Based Assay for the SH2 Domain of STAT4. Methods Protoc. 2022, 5, 93. https://doi.org/10.3390/mps5060093
Berg A, Gräber M, Schmutzler S, Hoffmann R, Berg T. A High-Throughput Fluorescence Polarization-Based Assay for the SH2 Domain of STAT4. Methods and Protocols. 2022; 5(6):93. https://doi.org/10.3390/mps5060093
Chicago/Turabian StyleBerg, Angela, Martin Gräber, Sebastian Schmutzler, Ralf Hoffmann, and Thorsten Berg. 2022. "A High-Throughput Fluorescence Polarization-Based Assay for the SH2 Domain of STAT4" Methods and Protocols 5, no. 6: 93. https://doi.org/10.3390/mps5060093