Recombinant Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
![Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-318503-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-western-blot.jpg "Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP799Y] to Calmodulin 1/2/3 - C-terminal
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, IHC-Fr
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal
See all Calmodulin 1/2/3 primary antibodies -
Description
Rabbit monoclonal [EP799Y] to Calmodulin 1/2/3 - C-terminal -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, IHC-Frmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Calmodulin 1/2/3 aa 100 to the C-terminus. The exact sequence is proprietary.
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Positive control
- WB: C6, RAW264.7, NIH/3T3, HeLa, HCT 116 cell lysate. IHC-P: Human urinary bladder carcinoma and testis tissues. Flow Cyt (intra): MCF7 and NIH/3T3 cells. IP: Human skeletal muscle tissue lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EP799Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab45689 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/50 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| WB | (9) |
1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 17 kDa).
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat. |
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| IP | (1) |
1/20 - 1/50.
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| IHC-Fr | (2) |
Use at an assay dependent concentration.
Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat. |
| Notes |
|---|
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Flow Cyt (Intra)
1/50 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB
1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 17 kDa). |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat. |
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IP
1/20 - 1/50. |
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IHC-Fr
Use at an assay dependent concentration. Mouse and rat species reactivity is recommended based on WB results, we do not guarantee IHC for mouse or rat. |
Target
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Relevance
Function: Calmodulin mediates the control of a large number of enzymes and other proteins by Ca(2+). Among the enzymes to be stimulated by the calmodulin-Ca(2+) complex are a number of protein kinases and phosphatases. Together with CEP110 and centrin, is involved in a genetic pathway that regulates the centrosome cycle and progression through cytokinesis. -
Cellular localization
Cytoplasm > cytoskeleton > spindle. Cytoplasm > cytoskeleton > spindle pole. Distributed throughout the cell during interphase, but during mitosis becomes dramatically localized to the spindle poles and the spindle microtubules. -
Database links
- Entrez Gene: 808 Human
- Entrez Gene: 12315 Mouse
- Entrez Gene: 24244 Rat
- Omim: 114182 Human
- Omim: 114183 Human
- SwissProt: 114180 Human
- SwissProt: P0DP23 Human
- SwissProt: P0DP24 Human
see all -
Form
There are three genes which encode an identical calcium binding protein which is one of the four subunits of phosphorylase kinase. -
Alternative names
- CALM 1 antibody
- CALM 2 antibody
- CALM 3 antibody
see all
Images
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Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)Burkard et al PLoS Pathog. 2017 Feb 23;13(2):e1006206. doi: 10.1371/journal.ppat.1006206. eCollection 2017 Feb. Fig 8. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
ΔSRCR peripheral blood monocyte-derived macrophages (PMMs) still function as hemoglobin-haptoglobin (Hb-Hp) scavengers.
Porcine PMMs were incubated for 24 hours in the presence of 100 μg/mol Hb-Hp then lysed with reducing SDS sample buffer and HO-1 protein expression analyzed by western blot.
For full details please see paper.
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![Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-266595-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-western-blot.jpg)
Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)All lanes : Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/10000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : HCT 116 (human colorectal carcinoma) whole cell lysate
Blocking peptides at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 17 kDaBlocking and Diluting buffer: 5% NFDM/TBST
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![Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-266600-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)ab45689 immunoprecipitating Calmodulin. 10 µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.
Lane 1: Human skeletal muscle lysate (10 µg)
Lane 2: Human skeletal muscle lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab45689 in human skeletal muscle lysate -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-266597-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)ab45689 staining Calmodulin in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control: PBS in place of primary antibody (inset).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-57813-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)ab45689 at a 1:250 dilution staining Calmodulin in human urinary bladder carcinoma tissue by immunohistochemistry in paraffin embedded tissue.
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![Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-266596-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-western-blot.jpg)
Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)All lanes : Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/10000 dilution
Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
Lane 2 : C6 (rat glioma) whole cell lysate
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: 17 kDaBlocking and diluting buffer: 5% NFDM/TBST
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![Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-57814-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-western-blot.jpg)
Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/5000 dilution + NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg
Predicted band size: 17 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted? -
![Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-57811-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)Overlay histogram showing MCF7 (Human breast adenocarcinoma cell line) cells stained with ab45689 (red line).
The cells were fixed with methanol (5 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 (ab45689, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C.
Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
This antibody gave a decreased signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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![Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-266599-anti-calmodulin-1-2-3-antibody-ep799y-c-terminal-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)ab45689 staining Calmodulin in NIH/3T3 (Mouse embryo fibroblast cell line) cells by intracellular flow cytometry.
Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluorr® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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![Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)](https://www.abcam.com/ps/products/45/ab45689/Images/ab45689-16-benefits-of-recombinant-antibodies.png)
Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
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
References (49)
ab45689 has been referenced in 49 publications.
- Sjøgaard-Frich LM et al. Dynamic Na+/H+ exchanger 1 (NHE1) - calmodulin complexes of varying stoichiometry and structure regulate Ca2+-dependent NHE1 activation. Elife 10:N/A (2021). PubMed: 33655882
- Nejedlá M et al. The actin regulator profilin 1 is functionally associated with the mammalian centrosome. Life Sci Alliance 4:N/A (2021). PubMed: 33184056
- Hook SC et al. TBC1D1 interacting proteins, VPS13A and VPS13C, regulate GLUT4 homeostasis in C2C12 myotubes. Sci Rep 10:17953 (2020). PubMed: 33087848
- Bae B et al. Elimination of Calm1 long 3'-UTR mRNA isoform by CRISPR-Cas9 gene editing impairs dorsal root ganglion development and hippocampal neuron activation in mice. RNA 26:1414-1430 (2020). PubMed: 32522888
- Chen P et al. Erianin, a novel dibenzyl compound in Dendrobium extract, inhibits lung cancer cell growth and migration via calcium/calmodulin-dependent ferroptosis. Signal Transduct Target Ther 5:51 (2020). PubMed: 32382060
