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Immunohistochemistry (PFA fixed) - CaMKII alpha (phospho T286) antibody (ab5683)
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Western blot - CaMKII alpha (phospho T286) antibody (ab5683)
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Western blot - CaMKII alpha (phospho T286) antibody (ab5683)
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Product Name
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CaMKII alpha (phospho T286) antibody
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See all CaMKII alpha antibodies (11)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to CaMKII alpha (phospho T286)
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Immunogen
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Synthetic phosphopeptide derived from a region of human CaM Kinase II alpha that contains threonine 286.
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Reacts with
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Hu, Rat, Rb
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Predicted to react
(species key)
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Ms
(Due to sequence homology)
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Tested applications
(see key)
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IHC (PFA fixed), IHC-Fr, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
IHC-Fr: Use at an assay dependent concentration.
Immunohistochemistry (PFA fixed): 1/300.
WB: 1/1000. Detects a band of approximately 50 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Rat brain lysate.
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Cellular localization
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Cytoplasmic; Cell membrane; presynaptic membrane. Cell junction; synapse. Postsynaptic lipid rafts.
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Research areas
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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
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Relevance
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Ca2+/CalmodulinDependent Protein Kinase II (CaM Kinase II) is a multifunctional calcium and calmodulindependent protein kinase that mediates cellular responses to a wide variety of intercellular signals. CaM Kinase II has been shown to regulate diverse cellular functions including synaptic plasticity, neurotransmitter synthesis and release, gene expression, ion channel function, carbohydrate metabolism, cytoskeletal function, and Ca2+homeostasis. Phosphorylation of Thr286 on the kinase produces an autonomously active form of CaM Kinase II. Autophosphorylation of Thr305 inhibits the activity CaM Kinase II. Phosphorylation at this site appears to reduce the association of CaM Kinase II with the PSD and reduce LTP and learning.
Calcium/calmodulin-dependent protein kinase II alpha (CaM Kinase II alpha) is a 50 kDa member of CaM Kinase II family of serine-threonine kinases that transduce Ca2+ signals to several target proteins, including ion channels and transcription activators. CaM Kinase II is predominantly expressed in two isoforms in the brain: alpha and beta. CaM Kinase II plays an important role in neuronal plasticity and memory formation, and exerts both calcium-calmodulin-dependent and -independent activities. Autophosphorylation of CaM Kinase II alpha on threonine 286 allows the kinase to switch from a calmodulin-dependent to a calmodulin-independent state, and is required for various cellular functions including hippocampal long-term potentiation (LTP), special learning, and hippocampus-dependent memory.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.05% Sodium Azide
Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3 Material safety datasheets (MSDS) for this product:
Glycerol MSDS
Sodium Azide MSDS
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Purification notes
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The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated CaM Kinase II alpha. The final product is generated by affinity chromatography using a CaM Kinase II alpha-derived peptide that is phosphorylated at threonine 286.
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this CaMKII alpha (phospho T286) antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for CaMKII alpha (phospho T286) antibody images, references, products related to ab5683 and other tools.
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CaMKII alpha (phospho T286) antibody images:
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 Immunohistochemistry (PFA fixed) - CaMKII alpha (phospho T286) antibody (ab5683)
Immunofluorescent staining for CaMKII alpha phospho (T286) using ab5683 (1/300, incubated for 18 hours) in rat spinal cord. To induce CaMKII alpha phospho (T286) protein expression, a noxious stimulus was administered to the rat 5 minutes prior to 4% PFA perfusion fixation (this is a known paradigm for inducing phosphorylation CaMKII in some spinal neurons). The resulting immunofluorescent staining for CaMKII alpha phospho (T286) protein is observed in the cytoplasm of many dorsal horn spinal neurons (surrounding the central canal [A] or in the ventral horn [B]). Omition of primary antibody resulted in a lack of staining (data not shown).
Protocol details: Tissue was prepared by 4% paraformaldehyde cardiac perfusion fixation. Tissue was frozen on dry ice and then embedded in OCT compound and cut on cryostat. An antigen retrieval step was not neccesary for the IHC protocol.
Sophie Pezet, Univ London Kings Coll, United Kingdom
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Western blot - CaMKII alpha (phospho T286) antibody (ab5683)
Peptide Competition and Phosphatase Treatment: Rat brain lysates were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda (ë) phosphatase (5) and blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with ab5683 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that only the peptide corresponding to CaM Kinase II alpha [pT286] blocks the antibody signal. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
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Western blot - CaMKII alpha (phospho T286) antibody (ab5683)
Primary Antibody 1/1000 24 hours at 4°C
Secondary Antibody (1/5000): HRP-conjugated Goat polyclonal to rabbit IgG
Blocking step: 5% BSA for 12 hours at 4°C This image is courtesy of an Abreview submitted by Dr. Byung-il Choi
See Abreview
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References for CaMKII alpha (phospho T286) antibody (ab5683)
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This product has been used in: (two most recent references)
Rafacho A et al. Glucocorticoids in vivo induce both insulin hypersecretion and enhanced glucose sensitivity of stimulus-secretion coupling in isolated rat islets. Endocrinology 151:85-95 (2010). WB; Rat. PubMed: 19880808
Tinsley CJ et al. A role for calcium-calmodulin-dependent protein kinase II in the consolidation of visual object recognition memory. Eur J Neurosci 30:1128-39 (2009). IHC-FoFr; Rat. PubMed: 19735285
See all 4 publication references for this product.
If you publish research using ab5683 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to CaMKII alpha
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