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| | LZK Biochemical Assay |
| Description: | Assay Protocol for 384 low volume MTP format: 1 μl of test compound was pipetted into test plate; +5 μl LZK solution (10 nM LZK, catalytic domain, Carna Biosiences in assay buffer); +5 μl substrate solution (15 nM MKK7, LDB; 15 μM ATP, in assay buffer); 90 min incubation at RT; +5 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 32° C.; endpoint measurement BMG (HTRF). All indicated concentrations final concentrations. Assay buffer (384 format): 50 mM Hepes pH 7.5, 10 mM MgCl2, 1 mM EGTA, 1 mM DTT, 0.01% Triton X-100, 0.01% BSA, 1:500 Smart Block. Detection buffer: 25 mM Tris/HCl pH 7.5, 100 mM EDTA, 100 mM NaCl, 200 mM KF, 0.01% Tween 20, 1:500 Smart Block. |
| Affinity data for this assay | |
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