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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 5 mM MOPS, 2.5 mM β-glycerophosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 0.05 mM DTT, 50 ?g/ml BSA, 2 mM ATP. 0.5 mM CaCl2, 0.03 mg/ml calmodulin. Incubate CaMK2α with 0.3 mg/ml autocamtide-2 peptide as substrate and 50 ?M [33P]-ATP at 30°C for 15 minutes, then spot reaction on phosphocellulose paper, fix in 1% phosphoric acid, and assay with a scintillation counter.
Background: CAMK2α is a ser/thr protein kinase that is a member of the Ca2+/calmodulin-dependent protein kinase family. CAMK2α is abundant in the brain as a major constituent of the postsynaptic density and is required for hippocampal long-term potentiation (LTP) and spatial learning. In addition to its Ca2+/calmodulin-dependent activity, CAMK2α can undergo autophosphorylation, resulting in Ca2+/calmodulin-independent activity. The protein level of CAMK2α fluctuates during neuronal activity in cultured rat pup hippocampal neurons. The levels of CAMK2α increased with heightened neuronal activity (2).
Description: Recombinant human calcium/calmodulin-dependent protein kinase II alpha (CaMK2α), GenBank Accession #NM_171825, full length, with an N-terminal GST-tag, MW = 74 kDa, expressed in Sf9 cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Genbank: NM_171825
Host Cell Line: Sf9 cells
Purity: ≥75%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q9UQM7
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Silva, A J. et a . Science 257: 206-211, 1992.
2. Thiagarajan, T C. et al . Neuron 36: 1103-1114, 2002.
3. Salas, M.A., et al. J. Mol. Cell Cardiol. (2010) 48(6):1298-130.
4. Chen, Y., et al. Neurochem Res. (2008) 33(10):2092-8.
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