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Neuroscience Neurotransmission Calcium Signaling Calmodulin / CaMK

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

  • Datasheet
  • SDS
Reviews (6)Q&A (4)References (38)

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Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
  • Immunocytochemistry/ Immunofluorescence - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
  • Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)

Key features and details

  • Rabbit polyclonal to CaMKII alpha (phospho T286)
  • Suitable for: IHC-FoFr, ICC/IF, WB
  • Reacts with: Rat
  • Isotype: IgG

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概述

  • 产品名称

    Anti-CaMKII alpha (phospho T286)抗体
    参阅全部 CaMKII alpha 一抗
  • 描述

    兔多克隆抗体to CaMKII alpha (phospho T286)
  • 宿主

    Rabbit
  • 特异性

    It does cross-react with CAMKII beta protein
  • 经测试应用

    适用于: IHC-FoFr, ICC/IF, WBmore details
  • 种属反应性

    与反应: Rat
  • 免疫原

    Synthetic peptide corresponding to CaMKII alpha (phospho T286).

  • 常规说明


    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.30
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.1% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 纯化说明

    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.
  • Primary antibody说明

    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.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Neuroscience
    • Neurotransmission
    • Calcium Signaling
    • Calmodulin / CaMK
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calmodulin Pathway
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Types of disease
    • Cancer
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human CaMKII alpha protein (ab60899)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab5683于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IHC-FoFr
1/300.
ICC/IF
Use a concentration of 5 µg/ml.
WB (4)
1/1000. Detects a band of approximately 50 kDa.
说明
IHC-FoFr
1/300.
ICC/IF
Use a concentration of 5 µg/ml.
WB
1/1000. Detects a band of approximately 50 kDa.

靶标

  • 功能

    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.
  • 序列相似性

    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
    Contains 1 protein kinase domain.
  • 细胞定位

    Cell junction > synapse > presynaptic cell membrane. Cell junction > synapse. Postsynaptic lipid rafts.
  • Target information above from: UniProt accession Q9UQM7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 25400 Rat
    • SwissProt: P11275 Rat
    • Unigene: 107499 Rat
    • 别名

      • 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

    图片

    • Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
      Western blot - Anti-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. Peptide Competiti
    • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
      Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-CaMKII alpha (phospho T286) antibody (ab5683)This image is courtesy of Sophie Pezet, Univ London Kings Coll, United Kingdom

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
      Immunocytochemistry/ Immunofluorescence - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
      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.

    • Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)
      Western blot - Anti-CaMKII alpha (phospho T286) antibody (ab5683)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

    实验方案

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (38)

    发表研究结果有使用 ab5683?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab5683 被引用在 38 文献中.

    • Jiang Y  et al. Aquaporin 1 mediates early responses to osmotic stimuli in endothelial cells via the calmodulin pathway. FEBS Open Bio 11:75-84 (2021). PubMed: 33125833
    • Wang XL  et al. Activation of a7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway. Aging (Albany NY) 12:543-570 (2020). PubMed: 31905173
    • Zhu X  et al. Rabies Virus Pseudotyped with CVS-N2C Glycoprotein as a Powerful Tool for Retrograde Neuronal Network Tracing. Neurosci Bull 36:202-216 (2020). PubMed: 31444652
    • Purohit DC  et al. Voluntary wheel running during adolescence distinctly alters running output in adulthood in male and female rats. Behav Brain Res 377:112235 (2020). PubMed: 31521739
    • He S  et al. Uncovering the Molecular Mechanism of the Qiang-Xin 1 Formula on Sepsis-Induced Cardiac Dysfunction Based on Systems Pharmacology. Oxid Med Cell Longev 2020:3815185 (2020). PubMed: 32908632
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-CaMKII alpha (phospho T286) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Feb 13 2014

    Western blot abreview for Anti-CaMKII alpha (phospho T286) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 04 2012

    Immunohistochemistry (Frozen sections) abreview for Anti-CaMKII alpha (phospho T286) antibody

    Poor
    Abreviews
    Abreviews
    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
    Read More

    MRS. Nicole Isaacs

    Verified customer

    提交于 May 10 2010

    Western blot abreview for Anti-CaMKII alpha (phospho T286) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Dr. Byung-il Choi

    Verified customer

    提交于 Feb 09 2010

    Western blot abreview for Anti-CaMKII alpha (phospho T286) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 04 2009

    Immunohistochemistry (Frozen sections) abreview for Anti-CaMKII alpha (phospho T286) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    DR. Sophie Pezet

    Verified customer

    提交于 Dec 19 2005

    Question

    Thanks very much for your reply. CaMK2 has several isoforms, including http://www.phosphosite.org/proteinAction.do?id=580, http://www.phosphosite.org/proteinAction.do?id=674, http://www.phosphosite.org/proteinAction.do?id=2093and http://www.phosphosite.org/proteinAction.do?id=2090. http://www.phosphosite.org/proteinAction.do?id=2093mainly exists in the heart. I want to know whether Anti-CaMK2 alpha (phospho T286) antibody (ab5683) can be used to examine phosphorilation of T286 of CaMK2-delta in rabbit heart (Western Blot). Or do you have another antibody that could be used to examine it?

    Another question: do you have anti-oxidized-CaMK2 antibody? I am looking forward to your answer.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2012

    Answer

    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, and the antibody should crossreact with the rabbit delta form. In any case, we would guarantee the antibody to work with rabbit samples.

    Unfortunately, we do not carry an antibody to oxidized CaMK2.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Jun 21 2012

    Question

    Now I need to detect CaMKII activation (pThr286-CaMKII) using rabbit cardiomyocytes. I want to know whether Anti-CaMKII alpha (phospho T286) antibody (ab5683) can react with rabbit heart.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 20 2012

    Answer

    Thank you for contacting us.

    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 valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise.


    I assume you will be using cultured cells or tissue sections. The following applications have been tested with this antibody: IHC-Frozen, ICC/IF (cultured cells), WB, IHC (PFA fixed).
    Paraffin embedded sections have not yet been tried.

    For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

    The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody/protein/kit at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody/protein at the full price (100%) of the antibody/protein you have tested, or a full price discount for the amount paid for the kit. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
    This programapplies to ELISA kits and other kits we offer.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Jun 20 2012

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 927009 DESCRIPTION OF THE PROBLEM no bands SAMPLE Rat hippocampus PRIMARY ANTIBODY Manufacturer: Abcam Inc. Species: Rabbit Diluent: 5% milk made in tTBS Dilution: tried 1:1000, 1:500, and 1:250 Incubation time: 1 hour Wash step: washed 3 times for 15 minutes in tTBS DETECTION METHOD Amersham ECL Western Blotting Detection Reagents (GE Healthcare) POSITIVE AND NEGATIVE CONTROLS USED After we tried to probe membranes with this antibody we reprobed them with three other antibodies and got strong signals. ANTIBODY STORAGE CONDITIONS The antibody was stored at -20 C and aliquoted after the first use. SAMPLE PREPARATION Buffer: Homogenization buffer: 600ul 5M NaCl (150mM) 400ul 1M Tris pH 7.5 (20mM) 80ul 0.5M EDTA (2mM) 200ul 0.1M DTT (1mM) 200ul Triton (1%) 40ul CLAP 18.48ml H2O Protease inhibitors: CLAP: Chymostatin Leupeptin Antipain Pepstatin Heating: 5min at 95 C before loading AMOUNT OF PROTEIN LOADED About 52ug. After we tried to probe membranes with this antibody we reprobed them with three other antibodies and got strong signals. ELECTROPHORESIS/GEL CONDITIONS Non-reducing, 10% Tris/Tricine Polyacrylamide Gel TRANSFER AND BLOCKING CONDITIONS Transfer buffer: 500ml 1X Tris/Tricine/SDS Running buffer 200ml Methanol 300ml dH2O Transfer time period: 50 minutes Blocking agent: 5% milk in tTBS for 1 hour SECONDARY ANTIBODY Manufacturer: Jackson Immuno Research Species: Goat Anti-Rabbit IgG Horseradish peroxidase Diluent: 5% milk made in tTBS Dilution: tried 1:10,000 Incubation time: 1 hour Wash step: washed 3 times for 15 minutes in tTBS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

    Read More

    Abcam community

    Verified customer

    Asked on May 18 2007

    Answer

    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 event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a credit note/refund will be offered. From the protocols questionnaire, I noticed that you used 5% milk as a blocking reagent and also as diluents for the primary and secondary antibodies. I believe that the reason why you are not seeing any bands is because cross reaction between the blocking agent and antibodies might have occurred. Here at Abcam, we recommend using 5% BSA as almost all of our products have been tested with it. Please try BSA as a substitute for milk. One other possible reason why no bands were observed could also be due to insufficient antibody binding to protein of interest. A longer incubation time will ensure proper and sufficient binding. Please try incubating the primary antibody at 4oC, overnight if you have not already done so. Can you confirm that the second antibody you used also managed to produce good results with other primary antibodies? I hope the above suggestions might prove to be useful in resolving the matter. Please try them out and should you still not get any results, please get back to me with your purchase details.

    Read More

    Abcam Scientific Support

    回复于 May 18 2007

    Question

    can this be used as a positive control in western blotting analysis of CaMKII alpha phosphorylated and non phosphorylated (ab5683 and ab3908 respectively).

    Read More

    Abcam community

    Verified customer

    Asked on Nov 07 2005

    Answer

    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-phospho pair)] also relates to these antibodies. This peptide pair can also be used to verify that the band you see is specific. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

    Read More

    Abcam Scientific Support

    回复于 Nov 08 2005

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