| Assay Method Information | |
| | LRRK2 Kinase Assay |
| Description: | The kinase assays for phosphorylation of LRRKtide (RLGRDKYKTLRQIRQ) or LRRKtideS (RLGRDKYKSLRQIRQ) were conducted in buffer containing 20 mM HEPES (pH 7.4), 50 mM NaCl, 10 mM MgCl2, 1 mM DTT, 0.5 mg/ml of BSA, 1 mM β-Gly-PO4, LRRKtide, ATP, and[γ-33P]ATP. The reactions were conducted in duplicate, initiated by the addition of 6 nM LRRK2, and incubated at room temperature for 150 min. The reactions were stopped by the addition of 20 mM EDTA and the mixture was transferred to a multiscreen PH filtration plate (Millipore, Billerica, MA) and washed six times with 75 mM H3PO4. The plate was dried, filters were removed, and the samples were counted with a scintillation counter. Background reactions were conducted in theabsence of LRRK2. |
| Affinity data for this assay | |
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