Recombinant Anti-NFAT1 antibody [EPR24658-149] - BSA and Azide free (ab283659)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24658-149] to NFAT1 - BSA and Azide free
- Suitable for: WB
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-NFAT1 antibody [EPR24658-149] - BSA and Azide free
See all NFAT1 primary antibodies -
Description
Rabbit monoclonal [EPR24658-149] to NFAT1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: ChIP,Flow Cyt (Intra),ICC/IF,IHC-P or IP -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Ramos, Raji, Daudi and EL4 whole cell lysates.
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General notes
ab283659 is the carrier-free version of ab283649.
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.
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. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24658-149 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283659 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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WB |
Use at an assay dependent concentration. Predicted molecular weight: 14 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 14 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
- Entrez Gene: 18019 Mouse
- Omim: 600490 Human
- SwissProt: Q13469 Human
- SwissProt: Q60591 Mouse
- Unigene: 713650 Human
- Unigene: 116802 Mouse
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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 [EPR24658-149] (ab283649) 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: 14 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?This data was developed using ab283649, the same antibody clone in a different buffer formulation.
False colour image of Western blot: Anti-NFAT1 antibody [EPR24658-149] 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, ab283649 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 [EPR24658-149] (ab283649) 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 : EL4 (mouse lymphoma T lymphocyte), whole cell lysate
Lane 5 : 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 (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 14 kDa
Observed band size: 100, 140 kDa why is the actual band size different from the predicted?This data was developed using ab283649, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
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: Lane 1, 2, 4, 5: 3 minutes ; Lane 3: 92 seconds
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 (0)
ab283659 has not yet been referenced specifically in any publications.