Anti-Calmodulin 1/2/3 antibody [2D1] (ab2860)
Key features and details
- Mouse monoclonal [2D1] to Calmodulin 1/2/3
- Suitable for: Flow Cyt, IHC-P, ICC
- Reacts with: Rat
- Isotype: IgG1
Overview
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Product name
Anti-Calmodulin 1/2/3 antibody [2D1]
See all Calmodulin 1/2/3 primary antibodies -
Description
Mouse monoclonal [2D1] to Calmodulin 1/2/3 -
Host species
Mouse -
Specificity
This antibody detects calmodulin. It does not detect parvalbumin, tropinin, S-100, or myosin light chain kinase (MLCK).By Western blot, this antibody detects a 17 kDa protein representing calmodulin from Dictyostelium cell lysate. Immunohistochemical staining of calmodulin in Dictyostelium cells with this antibody results in staining of the contractile vacuoles. -
Tested applications
Suitable for: Flow Cyt, IHC-P, ICCmore details -
Species reactivity
Reacts with: Rat -
Immunogen
Other Immunogen Type corresponding to Calmodulin 1/2/3. Calmodulin purified from Dictyostelium discoideum.
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Positive control
- ICC: HeLa, A2058, C6 cells. IHC-P: Rat testis and cerebellum tissue. Flow Cyt: C6, MCF-7 and PC-12 cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 99% PBS -
Concentration information loading...
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Purity
Protein A purified -
Primary antibody notes
Calmodulin is a small, highly conserved calcium binding protein found in all eukaryotic cells. With the capacity to bind up to four calcium ions, this 17 kDa protein acts as an important intracellular receptor for regulatory calcium signals. As it binds calcium, calmodulin undergoes conformational changes which can increase its affinity for target proteins. It acts both directly, through interaction with key target enzymes, and indirectly, via specific kinases. Studies have found that calmodulin participates in the regulation of several biological processes including energy and biosynthetic metabolism, cell motility, exocytosis, cytoskeletal assembly, and intracellular modulation of both cAMP and calcium concentrations. -
Clonality
Monoclonal -
Clone number
2D1 -
Isotype
IgG1 -
Research areas
Images
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Immunofluorescent analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Calmodulin with ab2860. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [2D1] (ab2860)
Immunohistochemistry was performed on normal biopsies of deparaffinized rat testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of C6 (Rat glial tumor cell line) cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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Immunofluorescent analysis of C6 (Rat glial tumor cell line) cells labeling Calmodulin ab2860. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [2D1] (ab2860)
Immunohistochemistry was performed on normal biopsies of deparaffinized rat cerebellum tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of PC-12 (Rat adrenal gland pheochromocytoma cell line) cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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Immunofluorescent analysis of A2058 (Human metastatic melanoma cell line) cells labeling Calmodulin ab2860. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
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Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of MCF7 (Human breast adenocarcinoma cell line) cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at a dilution of 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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Overlay histogram showing HeLa cells stained with ab2860 (red line). The cells were fixed with 80% 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. The cells were then incubated with the antibody (ab2860, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [2D1] (ab2860)
Immunolocalization of calmodulin in rat brain cells using ab2860 (1:100)