| Assay Method Information | |
| | In vitro LSD1 Inhibition Assay |
| Description: | Briefly, the compounds in DMSO were added into the LSD1 in the reaction buffer consisting of 50 mM Tris HCl, pH 7.5, and 1% DMSO, using Acoustic Technology (Echo 550) in nanoliter range and incubated for 15 min at room temperature. The reactions were initiated by adding peptide substrate [10 µM Histone H3 (1 21) K4me2 peptide (AnaSpec, San Jose, CA, USA)] to the reaction mixtures and incubated for 60 min at room temperature. The reactions were terminated by the addition of the detection mixture consisting final concentration of 0.1 Unit/mL horseradish peroxidase (Sigma, San Jose, CA, USA) and 10 µM Amplex UltraRed (Life Technologies, Grand Island, NY, USA) and fluorescence at Ex/Em = 535/590 nm was measured with kinetic mode of 5 min interval in envision for 30 min. |
| Affinity data for this assay | |
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