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Synthetic peptide corresponding to Rat CaMKII aa 281-294.
Database link: Q9UQM7
CaM kinase II, in a calcium and calmodulin dependent manner, phosphorylates many different brain substrates including synapsin I, tryptophan hydroxylase, tyrosine hydroxylase and nitric oxide synthase thereby performing regulatory functions associated with increases in intracellular free calcium. CaM kinase II is particularly abundant in the hippocampus and forebrain where it comprises ~1% of total protein. Within the neuron, CaM kinase II is a major component of the postsynaptic density fraction accounting for as much as 30% of total protein in the post synaptic density (PSD) region. Autophosphorylation of threonine-286 occurs after calcium/calmodulin activation and can enable CaM kinase II to remain active independent of free calcium. Autophosphorylation of CaM kinase II in hippocampal neurons, which results in relatively high levels of kinase activity even at basal calcium concentrations, may allow for both upward and downward regulation as opposed to simply on/off regulation.
Our Abpromise guarantee covers the use of ab2724 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||1/2000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).|
This image is courtesy of an anonymous abreview.
The blot was blocked with 5% BSA for 1 hour at room temperature.
ICC/IF image of ab2724 stained Hep cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2724, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.