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Assay Method Information |
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| | Gel-based ABPP Assay |
| Description: | Tissue and cell proteomes (50 μL) were treated with either FP-rhodamine (1 μM) or WHP01 (2 μM) for 30 min at 37 °C. The reactions were then quenched by addition of 4× SDS-PAGE loading buffer (20 μL). Competitive gel-based ABPP experiments were performed as previously described7. Samples were visualized in-gel using a ChemiDoc MP imaging system (Bio-Rad). The fluorescence from rhodamine is presented in gray scale. 2-5 s exposure times and 20-60 s exposure times were used for AIG1/ADTRP-transfected and native proteomes, respectively. |
| Affinity data for this assay | |
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