Recombinant
RabMAb

Recombinant Anti-CaMKII antibody [EPR6686(2)] (ab134041)

Rabbit recombinant monoclonal CaMKII antibody [EPR6686(2)]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 1 publication(s).

Overview

  • Product name

    Anti-CaMKII antibody [EPR6686(2)]
    See all CaMKII primary antibodies
  • Description

    Rabbit monoclonal [EPR6686(2)] to CaMKII
  • Host species

    Rabbit
  • Specificity

    The immunogen shares 100% homology with CaMK2 beta, 93% homology with CaMK2 gamma and 73% homology with CaMK2 alpha and delta. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human CaMKII aa 550-650 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • ICC/IF: Differentiated Neuro-2A. U87-MG cells. WB: HeLa whole cell lysate (ab150035), rat and mouse brain and human fetal brain tissue lysate IHC-P: Human brain tissue, Human clear cell cancer of kidney tissue FC: SH-SY5Y
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab134041 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 54 kDa.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Flow Cyt 1/10 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • All lanes : Anti-CaMKII antibody [EPR6686(2)] (ab134041) at 0.08 µg/ml (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Rat brain lysates
    Lane 3 : Mouse brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 54 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

    This antibody might cross-react with family members based on the high immunogen homology. That might be the reason why doublet are detected.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell cancer of kidney tissue sections labeling CaMKII with Purified ab134041 at 1:500 dilution (1.5 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
  • Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling CaMKII with purified ab134041 at 1:70 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • ab134041 staining CaMKII in Neuro-2A cells. The cells were differentiated with 20μM Trans-retinoic acid and serum starved (0.1%FBS/DMEM) for 48hours. They were then fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with purified ab134041 at a 5μg/ml concentration and ab78078, Mouse monoclonal [2G10] to beta III Tubulin, at 5μg/ml concentration, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green) and an anti-mouse AlexaFluor® 594 (ab150120) at 2 μg/ml (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

  • Overlay histogram showing SH-SY5Y cells stained with ab134041 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab134041, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • All lanes : Anti-CaMKII antibody [EPR6686(2)] (ab134041) at 1/1000 dilution (unpurified)

    Lane 1 : Human fetal brain lysate
    Lane 2 : HeLa lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-

    rabbit at 1/2000 dilution

    Predicted band size: 54 kDa

  • Immunohistochemical analysis of paraffin-embedded Human brain tissue labelling CaMKII with unpurified ab134041 at 1/250 dilution.

  • Immunofluorescent analysis of U87-MG cells labelling CaMKII with unpurified ab134041 at 1/250 dilution.

References

This product has been referenced in:

  • Zhang Y  et al. YiQiFuMai Powder Injection Attenuates Coronary Artery Ligation-Induced Heart Failure Through Improving Mitochondrial Function via Regulating ROS Generation and CaMKII Signaling Pathways. Front Pharmacol 10:381 (2019). Read more (PubMed: 31031629) »
  • Wu Q  et al. Nalmefene attenuates malignant potential in colorectal cancer cell via inhibition of opioid receptor. Acta Biochim Biophys Sin (Shanghai) 50:156-163 (2018). Read more (PubMed: 29267844) »
See all 3 Publications for this product

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