Mouse Anti-Calmodulin-Dependent Protein Kinase II (CAMKII)
CaMKII is an important member of calcium/calmodulinactivated protein kinase family, functioning in neural synaptic stimulation and T-cell receptor signaling. CaMKII is expressed in many different tissues but is specifically found in the neurons of the forebrain and its mRNA is found within the dendrites and the soma of the neuron. The CaMKII that is found in the neurons consist of two subunits of 52 (termed alpha genes) and 60kD (beta genes). CaMKII has catalytic and regulatory domains, as well as an ATP-binding domain, and a consensus phosphorylation site. The binding of Ca2+/calmodulin to its regulatory domain releases its auto inhibitory effect and activates the kinase. This kinase activation results in autophosphorylation at threonine 286. The threonine phosphorylation state of CaMKII can be regulated through PP1/PKA. Whereas PP1 (protein phosphatase 1) dephosphorylates phospho-CaMKII at Thr286, PKA (protein kinase A) prevents this dephosphorylation. Autophosphorylation also enables CaMKII to attain an enhanced affinity for NMDA receptors in postsynaptic densities.
Applications
Suitable for use in Immunofluorescence/Immunocytochemistry, Immunoprecipitation, Western Blot, Immunohistochemistry, RIA and ELISA. Other applications not tested.
Recommended Dilutions
Immunofluorescence (IC): 1:50 Western Blot: 0.1ug/ml. Detected CaMKII in 20ug of rat brain tissue extract by colorimetric Western Blot analysis using IgG (AP) goat anti-mouse as the secondary antibody. Immunohistochemistry: 1:2000 Optimal dilutions to be determined by the researcher.
Recommended Secondary Antibodies
I1904-04D: IgG, H&L (AP) Pab GtxMo I1904-06D: IgG, H&L, X-Adsorbed (AP) Pab GtxMo I1904-06J: IgG, H&L, X-Adsorbed (AP) Pab GtxMo I1904-25D: IgG, H&L (AP) Pab GtxMo
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.
Immunogen
Partially purified rat CaMKII.
Form
Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide, 50% glycerol.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes ~50-60kD protein, corresponding to the apparent molecular mass of rat CaMKII alpha subunit on SDS-PAGE immunblots. Both the phosphorylated and nonphosphorylated forms are recognized. Species Crossreactivity: human, mouse and bovine
