Anti-CaMKII (phospho T286)抗体(ab32678)

概述

  • 产品名称
    Anti-CaMKII (phospho T286)抗体
    参阅全部 CaMKII 一抗
  • 描述
    兔多克隆抗体to CaMKII (phospho T286)
  • 特异性
    This antibody is specific for the ~50 kDa alpha CaM Kinase II subunit and the ~60 kDa beta CaM Kinase II subunit phosphorylated at Thr286 in Western blots. Immunolabeling is blocked by the phosphopeptide used as the antigen but not by the corresponding dephosphopeptide.
  • 经测试应用
    适用于: WB, IHC-P, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Pig, Xenopus laevis
    预测可用于: Chicken, Cow
  • 免疫原

    Synthetic phosphopeptide corresponding to amino acids surrounding phospho-286 from Rat brain CaMKII.

  • 阳性对照
    • Rat cortex lysate.

性能

应用

Our Abpromise guarantee covers the use of ab32678 in the following tested applications.

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

应用 Ab评论 说明
WB 1/1000. Predicted molecular weight: 50 kDa. Predicted molecular weight: ~50 kDa for the alpha subunit and ~60 kDa for the beta subunit of CaMKII.
IHC-P Use a concentration of 2 µg/ml.
IHC-Fr Use at an assay dependent concentration.

靶标

Anti-CaMKII (phospho T286) antibody 图像

  • All lanes : Anti-CaMKII (phospho T286) antibody (ab32678) at 1/1000 dilution

    Lane 1 : Human myocardium tissue lysate, heart failure
    Lane 2 : Human myocardium tissue lysate, hypertrophy
    Lane 3 : Human myocardium tissue lysate, non-failing

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP conjugated sheep anti-rabbit at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa
    Observed band size : 50 kDa


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview.

    Blocked with 5% milk for 1 hour at RT.

    Incubated with primary antibody for 14 hours at 4°C in TBS-T20.

    See Abreview

  • ab32678 (2µg/ml) staining CaMKII (phospho T286) in human Brain:cortex:frontal-lateral using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclear/cytoplasmic compartments within the stellate cells and the myelinated fibres of white matter region .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.<

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa

    PC 12 cells were incubated at 37°C for 30 minutes with vehicle control (0 µM) and different concentrations of myricetin (ab120721). Increased expression of CaMKII (phospho T286) (ab32678) in PC 12 cells correlates with an increase in myricetin concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab32678at 1/500 dilution and ab52476 at 1/500 dilution overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

  • All lanes : Anti-CaMKII (phospho T286) antibody (ab32678) at 1/1000 dilution

    Lane 1 : Rat brain cortex lysate.
    Lane 2 : Rat brain cortex lysate preincubated with lambda-phosphatase.

    Lysates/proteins at 50 µg per lane.


    Predicted band size : 50 kDa
    Observed band size : 50,60 kDa (why is the actual band size different from the predicted?)
    Predicted molecular weight: ~50 kDa for the alpha subunit and ~60 kDa for the beta subunit of CaMKII.
    These images show the phosphospecificity of this antibody.
  • Anti-CaMKII (phospho T286) antibody (ab32678)参考文献

    This product has been referenced in:
    • Becerra R  et al. Reversible redox modifications of ryanodine receptor ameliorate ventricular arrhythmias in the ischemic-reperfused heart. Am J Physiol Heart Circ Physiol 311:H713-24 (2016). Read more (PubMed: 27422983) »
    • Yang Z  et al. Reverse of Acute and Chronic Morphine Tolerance by Lithocholic Acid via Down-Regulating UGT2B7. Front Pharmacol 7:404 (2016). WB ; Mouse . Read more (PubMed: 27847477) »

    See all 11 Publications for this product

    Product Wall

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (brain)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    15 µg
    Specification
    brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 19 2017

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (brain)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (gel 10%)
    Loading amount
    15 µg
    Specification
    brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 12 2017

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Pancreatic Cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% gradient)
    Loading amount
    20 µg
    Specification
    Pancreatic Cells
    Blocking step
    Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Mar 03 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Pancreas)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    No
    Specification
    Pancreas
    Blocking step
    Casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jan 31 2017

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Specification
    Heart
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Oct 26 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human Cardiomyocyte)
    Specification
    Human Cardiomyocyte
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Ethanol
    Username

    Abcam user community

    Verified customer

    提交于 Jun 18 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (Cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Loading amount
    20 µg
    Treatment
    ATX-II for 10 minutes
    Specification
    Cardiomyocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 15 2016

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (heart)
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Loading amount
    20 µg
    Specification
    heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 15 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Single Cell (Cardiomyocyte))
    Permeabilization
    No
    Specification
    Single Cell (Cardiomyocyte)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Fixative
    Ethanol
    Username

    Abcam user community

    Verified customer

    提交于 Jun 15 2016

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Spinal cord)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    Spinal cord
    Blocking step
    LiCOR blocking buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 24°C
    Username

    Maral Tajerian

    Verified customer

    提交于 Aug 27 2015

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