Anti-Calmodulin-Dependent Protein Kinase II (CAMKII)

CaMKII alpha, the alpha subunit of Ca2+calmodulin-dependent protein kinase II is part of a family of multifunctional protein kinases which play a major role in Ca2+-mediated signal transduction. CaMKIIalpha is expressed in many different tissues but is specifically found in the neurons of the forebrain and its mRNA is found within the dendrites as well as the soma of the neuron. The neuronal CaMKII consists of two major subunits of 52 and 60kD which are encoded by alpha-and beta-CaMKII genes, respectively. Additional isoforms are generated by alternative splicing of these as well as of the ubiquitious gama-and delta-CaMKII genes. Each subunit has an ATP-binding domain (arginine-X-X-serune/threonine), consensus phosphorylation site, catalytic domain, and a centrally located regulatory domain which has calmodulin binding activity (1-5). Activation and autophosphorylation of CaMKII may regulate numerous neuronal processes which includes two forms of synaptic plasticity, long term potentiation and long term depression (6,7). Neuronal CAMKII subunits assemble as large multimeric holoenzymes. The C-terminal association domains of 6-12 subunits assemble into a central globular structure from which the N-terminal catalytic/regulatory domains extend radially like petals of a flower (8). The subunit composition of the rat forebrain CaMKII holoenzyme consists of hetermers composed of alpha and beta subunits at a ratio of 2:1 and homomers composed of only alpha subunits. The association of CaMKII subunits leads to the positioning of their catalytic/regulatory domains in close proximity and the neighboring calmodulin-bound subunits cooperate to rapidly phosphorylate each other (9-11) Autophosphorylation also enables CaMKII to attain an enhanced affinity for NMDA receptors in postsynaptic densities (12-14), Applications: Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested. Recommended Dilutions: Western Blot (Colorimetric): 1:1000 , Immunohistochemistry: See reference (15)., Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.