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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is brain tissue.
CAMK2A (CaMKII alpha) is a serine/threonine kinase that regulates a number of cellular functions including neuronal processes, gene expression, carbohydrate metabolism, cytoskeletal function, and calcium homeostatsis. Autophosphorylation of threonine 286 in the presence of Ca2+ and calmodulin activates CAMK2A with the ability to switch from calmodulin-dependent to a calmodulin independent activity. CaMKII is an ubiquitously expressed serine/threonine protein kinase that is activated by Ca2+/calmodulin (CaM) binding and autophosphorylation and has been implicated in regulation of the cell cycle and transcription. It is involved in dendritic spine and synapse formation, neuronal plasticity and regulation of sarcoplasmic reticulum Ca2+ transport in skeletal muscle. In neurons, plays an essential structural role in the reorganization of the actin cytoskeleton during plasticity by binding and bundling actin filaments in a kinase-independent manner.
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