| Assay Method Information | |
| | DLK Biochemical Assay |
| Description: | Assay Protocol for 1536 MTP format: test compounds were predispensed (60 nl) in test plates; +3 μl DLK solution (10 nM DLK, catalytic domain, Carna Biosiences, 0.6 mM DTT in assay buffer); +2 μl substrate solution (15 nM MKK7, LDB; 60 μM ATP; 0.4 mM DTT in assay buffer); 90 min incubation at RT; +2 μl detection mix (5 nM XL665-labeled anti-His antibody, Cisbio; 0.1 nM Eu-labeled anti-phospho-MKK7 (T275/S277), Cisbio/Millipore in detection buffer); 16 h incubation at 37° C.; endpoint measurement BMG (HTRF). Assay Protocol for 384 low volume MTP format: 1 μl of test compounds were pipetted into test plate; +5 μl DLK solution (10 nM DLK, catalytic domain, Carna Biosiences in assay buffer); +5 μl substrate solution (15 nM MKK7, LDB; 60 μM ATP, in assay buffer); 90 min incubation at RT; +9 μl detection mix (20 nM XL665-labeled anti-His antibody, Cisbio; 0.4 nM Eu-labeled anti-phospho-MKK7 (T275/S277), Cisbio/Millipore in detection buffer); 16 h incubation at 4° C.; endpoint measurement BMG (HTRF). |
| Affinity data for this assay | |
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