| Assay Method Information | |
| | Biochemical Assay |
| Description: | Materials: LRRK2 G2019S enzyme Substrate (LRRKtide) ATP TR-FRET dilution buffer pLRRKtide antibody 384-well assay plate DMSOEnzyme Reaction Conditions 50 mM Tris pH 7.5, 10 mM MgCl2, 1 mM EGTA, 0.01% Brij-35, 2 mM DTT 5 nM LRRK2 134 μM ATP 60 minute reaction time 23° C. reaction temperature 10 μL total reaction volumeDetection Reaction Conditions 1× TR-FRET dilution buffer 10 mM EDTA 2 nM antibody 23° C. reaction temperature 10 μL total reaction volumeTo perform the detection of the reaction, EDTA completely mixed in TR-FRET dilution buffer was added to antibody reagent. 10 μL of detection reagent was added to all wells of each well of the assay plate and the plate was centrifuged to concentrate the mixture at the bottom of the wells. The plate was then incubated at 23° C. for 60 minutes. Plates were read on Perkin Elmer Envision 2104 instrument in TR-FRET mode using a 340 nm excitation filter, 520 nm fluorescence emission filter, and 490 or 495 nm terbium emission filter. |
| Affinity data for this assay | |
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