RabMAb

Anti-CaMKII抗体[EP1829Y] (ab52476)

概述

  • 产品名称Anti-CaMKII抗体[EP1829Y]
    参阅全部 CaMKII 一抗
  • 描述
    兔单克隆抗体[EP1829Y] to CaMKII
  • 特异性The peptide immunogen is highly conserved between CaMKII alpha, beta, gamma, and delta.
  • 经测试应用适用于: ICC/IF, WB, IHC-Pmore details
    不适用于: Flow Cyt or IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CaMKII aa 200-300.

  • 阳性对照
    • WB: Mouse brain, rat brain and human fetal brain tisse lysates. IHC-P: Human hepatocellulas carcinoma tissue. ICC/IF: U87-MG and PC12 cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

性能

应用

Our Abpromise guarantee covers the use of ab52476 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
ICC/IF 1/250.
WB 1/1000 - 1/5000. Detects a band of approximately 45, 70 kDa (predicted molecular weight: 54 kDa).

For unpurified use at 1/20000.

IHC-P 1/500.
  • 应用说明Is unsuitable for Flow Cyt or IP.
  • 靶标

    Anti-CaMKII antibody [EP1829Y] 图像

    • Anti-CaMKII antibody [EP1829Y] (ab52476) at 1/5000 dilution (purified) + Human fetal brain tissue lysate at 20 µg

      Secondary
      HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size : 54 kDa
      Observed band size : 45,70 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-CaMKII antibody [EP1829Y] (ab52476) at 1/1000 dilution (purified)

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size : 54 kDa
      Observed band size : 45,70 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-CaMKII antibody [EP1829Y] (ab52476) at 1/20000 dilution (unpurified) + Mouse brain lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP labeled at 1/2000 dilution

      Predicted band size : 54 kDa
      Observed band size : 45,70 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labelling CaMKII with purified ab52476 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling CaMKII with purified ab52476 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% aTriton X-100. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/250) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

    • ICC/IF image of unpurified ab52476 stained PC12 cells. The cells were 4% PFA 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 (ab52476, 1µg/ml) overnight at +4°C. The secondary antibody (green)ÿwas 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.

    Anti-CaMKII antibody [EP1829Y] (ab52476)参考文献

    This product has been referenced in:
    • Wang Y  et al. EPHB4 Protein Expression in Vascular Smooth Muscle Cells Regulates Their Contractility, and EPHB4 Deletion Leads to Hypotension in Mice. J Biol Chem 290:14235-44 (2015). Mouse . Read more (PubMed: 25903126) »
    • Cook-Snyder DR  et al. A retrograde adeno-associated virus for collecting ribosome-bound mRNA from anatomically defined projection neurons. Front Mol Neurosci 8:56 (2015). IHC (PFA fixed) ; Mouse . Read more (PubMed: 26557053) »

    See all 12 Publications for this product

    Product Wall

    Thanks so much for your reply.
    How is the tissue fixed and prepared before staining? If it is PFA fixed for more than ˜15 minutes, I'd suggest trying an antigen retrieval treatment in pH6 sodium citrate buffer, and a 1:100 dilution of primary anti...

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    Thank you very much for contacting us and letting us know about the trouble with this antibody.
    I do have a few questions, so that I can better understand the situation.
    1) How is the mouse brain tissue fixed and processed? Ab52476 has only b...

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    Thank you for contacting us.
    I am sorry to hear that the antibody is giving problems.

    As we discussed over the phone,I would suggest adding protease inhibitors to the lysis buffer in order to prevent the lysed proteins to be digested and...

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    I am beginning to gather the CoC's you have requested. When you have a chance, please provide the lot number from each catalog number so I can include this information on the documentation. Thanks for the additional information.

    Thank you for your enquiry. I can confirm that ab52476 CaMKII antibody [EP1829Y] recognizes all subunits of the protein, since the peptide immunogen is highly conserved between CaMKII-alpha, -beta, -gamma, and -omega. I hope this will be help...

    Read More
    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 cells expressing recombitant viral protein
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 minute(s) · Concentration: 0.5% · Temperature: 20°C
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    提交于 Aug 04 2009

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