Recombinant Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1670(2)] to Calcineurin A
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Calcineurin A antibody [EPR1670(2)]
See all Calcineurin A primary antibodies -
Description
Rabbit monoclonal [EPR1670(2)] to Calcineurin A -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
Unsuitable for: IHC-P or 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
- Fetal brain, SH-SY5Y, A431, and HeLa cell lysates; HeLa cells. Mouse and rat brain cortex lysates.
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1670(2) -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109412 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) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/10000 - 1/50000. Detects a band of approximately 58 kDa (predicted molecular weight: 59 kDa).
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ICC/IF |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 58 kDa (predicted molecular weight: 59 kDa). |
ICC/IF
1/100 - 1/250. |
Target
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Function
Calcium-dependent, calmodulin-stimulated protein phosphatase. This subunit may have a role in the calmodulin activation of calcineurin. Dephosphorylates DNM1L, HSPB1 and SSH1. -
Sequence similarities
Belongs to the PPP phosphatase family. PP-2B subfamily. -
Cellular localization
Nucleus. Colocalizes with ACTN1 and MYOZ2 at the Z line in heart and skeletal muscle. - Information by UniProt
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Database links
- Entrez Gene: 5530 Human
- Entrez Gene: 19055 Mouse
- Entrez Gene: 24674 Rat
- Omim: 114105 Human
- SwissProt: Q08209 Human
- SwissProt: P63328 Mouse
- SwissProt: P63329 Rat
- Unigene: 435512 Human
see all -
Alternative names
- Alpha isoform formerly PPP2B antibody
- Calcineurin A alpha antibody
- Calcineurin A1 antibody
see all
Images
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All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/10000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PPP3CA knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 59 kDa
Observed band size: 59 kDaLanes 1- 2: Merged signal (red and green). Green - ab109412 observed at 59 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109412 was shown to react with Calcineurin A in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265130 (knockout cell lysate ab257181) was used. Wild-type HeLa and PPP3CA knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109412 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent staining of HeLa cells using 1/100 ab109412.
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Overlay histogram showing HeLa cells stained with ab109412 (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 followed by the antibody (ab109412, 1/100 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) (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.
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All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/3000 dilution
Lane 1 : Mouse brain cortex lysate
Lane 2 : Mouse lung lysate
Lane 3 : Rat brain cortex lysate
Lane 4 : Rat liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 59 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted? -
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Calcineurin A knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab109412 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109412 was shown to specifically react with Calcineurin A when Calcineurin A knockout samples were used. Wild-type and Calcineurin A knockout samples were subjected to SDS-PAGE. ab109412 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/10000 dilution
Lane 1 : fetal brain lysate
Lane 2 : SH-SY5Y lysate
Lane 3 : A431 lysate
Lane 4 : HeLa cells lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 59 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab109412 has been referenced in 5 publications.
- Li L et al. Calf Thymus Polypeptide Restrains the Growth of Colorectal Tumor via Regulating the Intestinal Microbiota-Mediated Immune Function. Front Pharmacol 13:898906 (2022). PubMed: 35662701
- Moon JH et al. Calcineurin Activation by Prion Protein Induces Neurotoxicity via Mitochondrial Reactive Oxygen Species. Oxid Med Cell Longev 2021:5572129 (2021). PubMed: 34394828
- Gu LF et al. Huangkui Capsule Ameliorates Renal Fibrosis in a Unilateral Ureteral Obstruction Mouse Model Through TRPC6 Dependent Signaling Pathways. Front Pharmacol 11:996 (2020). PubMed: 32719603
- Liu Z et al. Discovery of Small-Molecule Inhibitors of the HSP90-Calcineurin-NFAT Pathway against Glioblastoma. Cell Chem Biol 26:352-365.e7 (2019). PubMed: 30639261
- Tian CJ & Pang X Ca2+-calcineurin signaling is involved in norepinephrine-induced cardiac fibroblasts activation. Int J Clin Exp Pathol 8:5210-6 (2015). PubMed: 26191219