Recombinant Anti-CaMKII antibody [EPR6686(2)] - BSA and Azide free (ab227108)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6686(2)] to CaMKII - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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 -
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.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Human fetal brain and HeLa lysates; Human brain tissue; U87-MG cells. ICC/IF: Differentiated Neuro-2A.
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General notes
ab227108 is the carrier-free version of ab134041.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6686(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
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. |
Target
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Function
CaM-kinase II (CAMK2) is a prominent kinase in the central nervous system that may function in long-term potentiation and neurotransmitter release. Member of the NMDAR signaling complex in excitatory synapses it may regulate NMDAR-dependent potentiation of the AMPAR and synaptic plasticity. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
Contains 1 protein kinase domain. -
Cellular localization
Cell junction > synapse > presynaptic cell membrane. Cell junction > synapse. Postsynaptic lipid rafts. - Information by UniProt
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Database links
- Entrez Gene: 815 Human
- Entrez Gene: 816 Human
- Entrez Gene: 817 Human
- Entrez Gene: 818 Human
- Entrez Gene: 108058 Mouse
- Entrez Gene: 12322 Mouse
- Entrez Gene: 12323 Mouse
- Entrez Gene: 12325 Mouse
see all -
Alternative names
- Calcium/calmodulin dependent protein kinase II alpha antibody
- Calcium/calmodulin dependent protein kinase II beta antibody
- Calcium/calmodulin dependent protein kinase II delta antibody
see all
Images
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Intracellular 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).
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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).
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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).
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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).
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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.
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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).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab227108 has been referenced in 1 publication.
- 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 . PubMed: 28443064