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) »
  • Ljubojevic S  et al. Early remodeling of perinuclear Ca2+ stores and nucleoplasmic Ca2+ signaling during the development of hypertrophy and heart failure. Circulation 130:244-55 (2014). Human . Read more (PubMed: 24928680) »

See all 9 Publications for this product

Product Wall

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

Dr. maral tajerian

Verified customer

提交于 Aug 27 2015


J’ai contacté le laboratoire qui m’ont confirmé qu’il ne détermine pas la concentration pour chaque lot mais qu’en général la concentration et de 0.25 mg/ml.
Comme expliqué au t...

Read More
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample Human Cell (Cardiomyocytes)
Specification Cardiomyocytes
Permeabilization No
Fixative Methanol
Username

Abcam user community

Verified customer

提交于 Feb 09 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample Mouse Cell (Cardiomyocytes)
Specification Cardiomyocytes
Permeabilization No
Fixative Ethanol
Username

Abcam user community

Verified customer

提交于 Jan 29 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Human Tissue lysate - whole (Right Atrium)
Specification Right Atrium
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Jan 29 2015

1-10 of 18 Abreviews or Q&A

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