Recombinant Anti-NFAT1 antibody [EPR24658-43] (ab283691)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24658-43] to NFAT1
- Suitable for: Flow Cyt (Intra), IP, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NFAT1 antibody [EPR24658-43]
See all NFAT1 primary antibodies -
Description
Rabbit monoclonal [EPR24658-43] to NFAT1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, WBmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Ramos, Raji, and Daudi whole cell lysates Flow cyt-intra: Ramos and Jurkat cells. IP: Ramos whole cell 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.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24658-43 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283691 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) |
1/50.
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IP |
1/1000.
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WB |
1/1000. Predicted molecular weight: 100 kDa.
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Notes |
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Flow Cyt (Intra)
1/50. |
IP
1/1000. |
WB
1/1000. Predicted molecular weight: 100 kDa. |
Target
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Function
Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2, IL-3, IL-4, TNF-alpha or GM-CSF. -
Tissue specificity
Expressed in thymus, spleen, heart, testis, brain, placenta, muscle and pancreas. -
Sequence similarities
Contains 1 RHD (Rel-like) domain. -
Domain
Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors. -
Post-translational
modificationsIn resting cells, phosphorylated by NFATC-kinase on at least 18 sites in the 99-363 region. Upon cell stimulation, all these sites except Ser-243 are dephosphorylated by calcineurin. Dephosphorylation induces a conformational change that simultaneously exposes an NLS and masks an NES, which results in nuclear localization. Simultaneously, Ser-53 or Ser-56 is phosphorylated; which is required for full transcriptional activity. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasmic for the phosphorylated form and nuclear after activation that is controlled by calcineurin-mediated dephosphorylation. Rapid nuclear exit of NFATC is thought to be one mechanism by which cells distinguish between sustained and transient calcium signals. The subcellular localization of NFATC plays a key role in the regulation of gene transcription. - Information by UniProt
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Database links
- Entrez Gene: 4773 Human
- Omim: 600490 Human
- SwissProt: Q13469 Human
- Unigene: 713650 Human
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Alternative names
- AI607462 antibody
- cytoplasmic 2 antibody
- KIAA0611 antibody
see all
Images
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All lanes : Anti-NFAT1 antibody [EPR24658-43] (ab283691) at 1/1000 dilution
Lane 1 : Wild-type Raji cell lysate
Lane 2 : NFATC2 CRISPR-Cas9 edited Raji cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 100 kDa
Observed band size: 100 kDaFalse colour image of Western blot: Anti-NFAT1 antibody [EPR24658-43] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283691 was shown to bind specifically to NFAT1. A band was observed at 100 kDa in wild-type Raji cell lysates with no signal observed at this size in NFATC2 CRISPR-Cas9 edited cell line ab280906 (CRISPR-Cas9 edited cell lysate ab282940). The band observed in the CRISPR-Cas9 edited lysate lane below 100 kDa is likely to represent a truncated form of NFAT1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and NFATC2 CRISPR-Cas9 edited Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-NFAT1 antibody [EPR24658-43] (ab283691) at 1/1000 dilution
Lane 1 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 2 : Raji (human Burkitts lymphoma B lymphocyte), whole cell lysate
Lane 3 : Daudi (human Burkitts lymphoma lymphoblast), whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 100 kDaBlocking and diluting buffer and concentration: 5% NFDM/TBSTThis blot was developed using a higher-sensitivity ECL substrate.
The molecular weight observed is consistent with what has been described in the literature (PMID:21078663, PMID:25696812).
Negative control: Hela (PMID:21078663)
Exposure time: 3 minutes
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell, Left) / Ramos (Human Burkitt's lymphoma B lymphocyte, Right) cells labelling NFAT1 with ab283691 at 1/50 dilution (1ug)/ red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: Hela (PMID:21078663).
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (Human prostate carcinoma epithelial cell, Left) / Jurkat (Human T cell leukemia T lymphocyte, Right) cells labelling NFAT1 with ab283691 at 1/50 dilution (1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: LNCaP.
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NFAT1 was immunoprecipitated from Ramos (Human Burkitt's lymphoma B lymphocyte), whole cell lysate with ab283691 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283691 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1 (Input): Ramos (Human Burkitt's lymphoma B lymphocyte), whole cell lysate, 10 μg
Lane 2 (+): Ramos whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab283691 in Ramos whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab283691 has not yet been referenced specifically in any publications.