![Immunocytochemistry/ Immunofluorescence - CaMKII alpha (phospho T286) antibody [22B1] (ab2724) image](#CaMKII-alpha-Primary-antibodies-ab2724-2.jpg)
![Flow Cytometry - Anti-CaMKII alpha (phospho T286) antibody [22B1] (ab2724) image](#CaMKII-alpha-Primary-antibodies-ab2724-3.jpg)
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Products:Neuroscience >> Neurotransmission >> Calcium Signaling >> Calmodulin / CaMK
Overview
Product name
Anti-CaMKII alpha (phospho T286) antibody [22B1]
See all CaMKII alpha products (11) ...
Description
Mouse monoclonal [22B1] to CaMKII alpha (phospho T286)
Specificity
Detects phosphorylated CaM kinase II from rat tissues. This antibody has been shown to detect the phosphorylated alpha subunit CaM kinase II only. It does not detect the non-phosphorylated alpha subunit CaM kinase II nor the 60 kDa beta subunit in either phosphorylation state. Immunofluorescence staining of CaM kinase II in rat hippocampal cells with this antibody densely labels cell somas and neuropil areas, while nuclei are only lightly stained.
Tested applications
Flow Cyt, ICC/IF, ELISA, IP, WBmore details
Cross reactivity
Reacts with
Rat, Human
Immunogen
Synthetic peptide: (281)MHRQET(p)VDCLKKFN(294). Immunizing peptide corresponds to amino acid residues 281-294 of rat CaM kinase II with the threonine at position 286 being phosphorylated.
MHRQETpVDC LKKFN
Positive control
WB: rat brain homogenate ICC: rat hippocampal cells in culture
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
IgG fraction
Primary antibody notes
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.
Clonality
Monoclonal
Clone number
22B1
Isotype
IgG1
Research areas
Cancer >> Cancer Metabolism >> Metabolic signaling pathway >> Metabolism of carbohydrates
Cardiovascular >> Heart >> Hypertrophy >> Other
Signal Transduction >> Signaling Pathway >> Calcium Signaling >> Calmodulin Pathway
Neuroscience >> Neurotransmission >> Calcium Signaling >> Calmodulin / CaMK
Applications
Show applications key
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.
ICC/IF: 1/100
ELISA: Use at an assay dependent dilution.
IP: Use at an assay dependent dilution.
WB: 1/2000Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).
Target
Function
CaM-kinase II (CAMK2) is a prominent kinase in the central nervous system that may function in long-term potentiation and neurotransmitter release. Member of the NMDAR signaling complex in excitatory synapses it may regulate NMDAR-dependent potentiation of the AMPAR and synaptic plasticity.
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
Contains 1 protein kinase domain.
Cellular localization
Cell junction > synapse > presynaptic cell membrane. Cell junction > synapse. Postsynaptic lipid rafts.
Target information above from: UniProt accessionQ9UQM7
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- alpha CaMKII antibody
- alpha CaMKII antibody
- Calcium / Calmodulin Dependent Protein Kinase II G antibody
see all
Database links
- Entrez Gene: 815 Human
- Entrez Gene: 25400 Rat
- Omim: 114078 Human
- SwissProt: Q9UQM7 Human
- SwissProt: P11275 Rat
- Unigene: 725992 Human
- Unigene: 107499 Rat
Anti-CaMKII alpha (phospho T286) antibody [22B1] images:
Western blot
Lane 1: unphosphorylated rat brain homogenate - ab2724
Lane 2: phosphorylated rat brain homogenate - ab2724
Lane 3: unphosphorylated rat brain homogenate - ab2725
Lane 4: phosphorylated rat brain homogenate - ab2725
Immunocytochemistry/ Immunofluorescence - CaMKII alpha (phospho T286) antibody [22B1] (ab2724)
![Immunocytochemistry/ Immunofluorescence - CaMKII alpha (phospho T286) antibody [22B1] (ab2724)](/ps/datasheet/images/2/ab2724/CaMKII-alpha-Primary-antibodies-ab2724-2.jpg)
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.
Flow Cytometry - Anti-CaMKII alpha (phospho T286) antibody [22B1] (ab2724)
![Flow Cytometry - Anti-CaMKII alpha (phospho T286) antibody [22B1] (ab2724)](/ps/datasheet/images/2/ab2724/CaMKII-alpha-Primary-antibodies-ab2724-3.jpg)
Overlay histogram showing SH-SY5Y cells stained with ab2724 (red line). The cells were fixed with 4% paraformaldehyde and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2724, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
References for Anti-CaMKII alpha (phospho T286) antibody [22B1] (ab2724)
This product has been referenced in:
- Jung SCet al. Rapid, bidirectional remodeling of synaptic NMDA receptor subunit composition by A-type K+ channel activity in hippocampal CA1 pyramidal neurons. Neuron 60:657-71 (2008). ICC/IF; Rat.Read more (PubMed: 19038222) »
- Trinidad JCet al. Comprehensive identification of phosphorylation sites in postsynaptic density preparations. Mol Cell Proteomics 5:914-22 (2006). WB; Mouse.Read more (PubMed: 16452087) »
See all 2 publications for this product
Publishing research using ab2724? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"