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Assay Method Information |
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| | Biophysical Assays |
| Description: | Isothermal titration calorimetry (ITC) was performed at 25 °C in a Nano ITC instrument (TA Instruments, USA). The cell was loaded with 25 μM PRMT6, 100 μM SAMand 2.5% DMSO in 50 mM Tris-HCl (pH 8.5) containing 150 mM NaCl. The syringe wasloaded with 200 μM MS023, 100 μM SAM and 2.5% DMSO prepared in the same buffer. 2 μL volumes were injected at 180 seconds intervals (25 injections in total). The data was analyzed using "Nano Analyze" software supplied by the manufacturer and fit to a one binding site model. Differential scanning fluorimetry was carried out as described previously.6 The experiments were done with 100 mM HEPES buffer (pH 7.4) containing 150 mM NaCl, 0.1 mg mL-1 PRMT6, and 5X Sypro Orange. |
| Affinity data for this assay | |
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