Recombinant
RabMAb

Recombinant Anti-CaMKII antibody [EPR6686(2)] - BSA and Azide free (ab227108)

Overview

  • Product name

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

    Rabbit monoclonal [EPR6686(2)] to CaMKII - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, WBmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CaMKII aa 550-650 (internal sequence).

  • Positive control

    • Human fetal brain and HeLa lysates; Human brain tissue; U87-MG cells. ICC/IF: Differentiated Neuro-2A.
  • General notes

    Ab227108 is the carrier-free version of ab134041. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227108 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227108 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
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
  • Application notes
    Is unsuitable for IP.
  • Target

    Images

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

    • 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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

    • 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).

       

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134041).

    • This ICC/IF data was generated using the same anti-CaMKII antibody clone, EPR6686(2), in a different buffer formulation (cat# ab134041).

      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 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).

       

    References

    This product has been referenced in:

    • Ndjim M  et al. Loss of Vagal Sensitivity to Cholecystokinin in Rats Born with Intrauterine Growth Retardation and Consequence on Food Intake. Front Endocrinol (Lausanne) 8:65 (2017). WB ; Rat . Read more (PubMed: 28443064) »
    See 1 Publication for this product

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