製品の概要

  • 製品名
    Anti-CaMKII (phospho T286) antibody
    CaMKII 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to CaMKII (phospho T286)
  • 特異性
    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.
  • アプリケーション
    適用あり: WB, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Pig, Xenopus laevis
    交差が予測される動物種: Chicken, Cow
  • 免疫原

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

  • ポジティブ・コントロール
    • Rat cortex lysate.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

Anti-CaMKII (phospho T286) antibody 画像

  • 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) 使用論文

This product has been referenced in:
  • 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) »
  • 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) »

See all 16 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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

投稿 Aug 27 2015

1-10 of 22 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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