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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Varying amounts of enzyme were incubated in 5 mM MOPS, pH 7.2, 2.5 mM β-glycero-phosphate, 4 mM MgCl2, 2.5 mM MnCl2, 1 mM EGTA, 0.4 mM EDTA, 0.01 mM DTT, 0.05 mM ATP, 5 µg of autocamtide 2 synthetic peptide substrate, and 4 µCi of [33P]-ATP was incubated at 30°C for 15 minutes. Then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: DCAMKL1 or doublecortin-like kinase 1 contains two N-terminal doublecortin domains (which bind microtubules and regulate microtubule polymerization), a C-terminal serine/threonine protein kinase domain (which shows substantial homology to Ca2+/calmodulin-dependent protein kinase), and a serine/proline-rich domain in between the doublecortin and the protein kinase domains (which mediates multiple protein-protein interactions) (1). DCAMKL1 is a microtubule-associated kinase that can undergo autophosphorylation. DCAMKL1 has microtubule-polymerizing activity that is independent of its protein kinase activity (2). DCAMKL1 is involved in several different cellular processes, including neuronal migration, retrograde transport, neuronal apoptosis and neurogenesis.
Description: Recombinant humanDCAMKL1, also known as DCLK1,DCDC3A, DCLK, or KIAA0369, GenBankAccession No. NM_004734, full-lengthwith an N-terminal GST tag andexpressed in a baculovirus-infected Sf9cell expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol.
Genbank: NM_004734
Host Cell Line: Sf9 cells
Purity: ≥75%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: O15075
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Ohmae, S. et.al: Molecular identification and characterization of a family of kinases with homology to Ca(2+)/calmodulin-dependent protein kinases I/IV. J. Biol. Chem. 281: 20427-20439, 2006.
2.Lin, P. T. et,al: DCAMKL1 encodes a protein kinase with homology to doublecortin that regulates microtubule polymerization. J. Neurosci. 20: 9152-9161, 2000.
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