Anti-CaMKII alpha (phospho T286) antibody (ab5683)

Overview

  • Product nameAnti-CaMKII alpha (phospho T286) antibody
    See all CaMKII alpha primary antibodies
  • Description
    Rabbit polyclonal to CaMKII alpha (phospho T286)
  • SpecificityIt does cross-react with CAMKII beta protein
  • Tested applicationsSuitable for: IHC-Fr, ICC/IF, WB, IHC (PFA fixed)more details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
  • Immunogen

    Synthetic phosphopeptide derived from a region of human CaM Kinase II alpha that contains threonine 286.

  • Positive control
    • Rat brain lysate.
  • General notes


    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.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesThe 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.
  • Primary antibody notesCalcium/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.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5683 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
WB 1/1000. Detects a band of approximately 50 kDa.
IHC (PFA fixed) 1/300.

Target

  • FunctionCaM-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 similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
    Contains 1 protein kinase domain.
  • Cellular localizationCell junction > synapse > presynaptic cell membrane. Cell junction > synapse. Postsynaptic lipid rafts.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha CaMKII antibody
    • Calcium calmodulin dependent protein kinase II antibody
    • Calcium/calmodulin dependent protein kinase II alpha B subunit antibody
    • Calcium/calmodulin dependent protein kinase type II alpha chain antibody
    • Calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha antibody
    • Calcium/calmodulin-dependent protein kinase II alpha antibody
    • Calcium/calmodulin-dependent protein kinase II-alpha antibody
    • Calcium/calmodulin-dependent protein kinase type II subunit alpha antibody
    • Calcium/calmodulin-dependent protein kinase type IIA antibody
    • CaM kinase II alpha chain antibody
    • CaM kinase II alpha subunit antibody
    • CaM kinase II subunit alpha antibody
    • CaMK II alpha subunit antibody
    • CaMK-II subunit alpha antibody
    • Camk2a antibody
    • CAMKA antibody
    • CaMKII antibody
    • CaMKIINalpha antibody
    • EC 2.7.11.17 antibody
    • KCC2A_HUMAN antibody
    • KIAA0968 antibody
    • MGC123320 antibody
    • MGC139375 antibody
    • MGC155201 antibody
    • mKIAA0968 antibody
    • PK2CDD antibody
    • PKCCD antibody
    • R74975 antibody
    • zgc:112538 antibody
    • zgc:123320 antibody
    see all

Anti-CaMKII alpha (phospho T286) antibody images

  • 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.

  • ICC/IF image of ab5683 stained PC12 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 (ab5683, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  • 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. Peptide Competiti
  • This image is courtesy of an Abreview submitted by Dr. Byung-il Choi

    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

    See Abreview

References for Anti-CaMKII alpha (phospho T286) antibody (ab5683)

This product has been referenced in:
  • Oka Y  et al. Thirst driving and suppressing signals encoded by distinct neural populations in the brain. Nature 520:349-52 (2015). IHC (PFA fixed) ; Mouse . Read more (PubMed: 25624099) »
  • Zhong W  et al. Serotonin 5-HT3 receptor-mediated vomiting occurs via the activation of Ca2+/CaMKII-dependent ERK1/2 signaling in the least shrew (Cryptotis parva). PLoS One 9:e104718 (2014). IHC-Fr . Read more (PubMed: 25121483) »

See all 11 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10% separating gel)
Sample Mouse Tissue lysate - whole (Hippocampal homogenate)
Specification Hippocampal homogenate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 13 2014

Thank you for your email.

For ab5683: I have checked the homology between the human CaMK2 alpha and rabbit CaMK2 delta around the phosphorylation site T286: It is 90-100% with maximal one residue mismatching. Thus the homology is very high, ...

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The antibody has been tested with rabbit samples and would be guaranteed for this species.

We guarantee all our products for TESTED species and applications as listed on the datasheet. The guarantee is ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (EL4)
Loading amount 15 µg
Specification EL4
Treatment cells expressing recombitant viral protein
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 04 2012

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Duodenum Biopsies)
Specification Duodenum Biopsies
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Mrs. Nicole Isaacs

Verified customer

Submitted May 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rabbit Cell lysate - whole cell (mouse hippocampal lysate)
Loading amount 5 µg
Specification mouse hippocampal lysate
Gel Running Conditions Reduced Denaturing (10%)
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. Byung-il Choi

Verified customer

Submitted Feb 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (huh-7)
Loading amount 15000 cells
Specification huh-7
Treatment recombitant viral protein expressed by cell
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 04 2009

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab5683 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the e...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Sections of rat spinal cord and brain)
Specification Sections of rat spinal cord and brain
Fixative Paraformaldehyde
Username

Dr. Sophie Pezet

Verified customer

Submitted Dec 19 2005

Thank you for your enquiry. Yes, ab7942 (rat brain lysate) or ab7188 (mouse brain lysate) could be used as a positive control for ab3908 and ab5683. The previous email that I sent about ab10772 [CaMKII alpha peptide - phospho T286 (phospho and non-...

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