Overview

  • Product name

    Anti-NFAT4/NF-ATc3 antibody
    See all NFAT4/NF-ATc3 primary antibodies
  • Description

    Rabbit polyclonal to NFAT4/NF-ATc3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cynomolgus monkey
  • Immunogen

    Synthetic peptide corresponding to Human NFAT4/NF-ATc3 aa 1000 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab102063)

  • Positive control

    • Recombinant Human NFAT4/NF-ATc3 protein (ab114667) can be used as a positive control in WB. This antibody gave a positive signal in human thymus and human kidney tissue lysates and in the following cell lines: Jurkat; Ramos; Raji. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal kidney. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: Jurkat.
  • General notes

    Previously labelled as NFAT4.

Properties

Applications

Our Abpromise guarantee covers the use of ab93628 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 116 kDa (predicted molecular weight: 116 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2.
  • Tissue specificity

    Isoform 1 is predominantly expressed in thymus and is also found in peripheral blood leukocytes and kidney. Isoform 2 is predominantly expressed in skeletal muscle and is also found in thymus, kidney, testis, spleen, prostate, ovary, small intestine, heart, placenta and pancreas. Isoform 3 is expressed in thymus and kidney. Isoform 4 is expressed in thymus and skeletal muscle.
  • Sequence similarities

    Contains 1 RHD (Rel-like) domain.
  • Domain

    Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
  • Post-translational
    modifications

    Phosphorylated by NFATC-kinase; dephosphorylated by calcineurin.
  • 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
  • Database links

  • Alternative names

    • C80703 antibody
    • cytoplasmic 3 antibody
    • D8Ertd281e antibody
    • NF AT4 antibody
    • NF ATc3 antibody
    • NF-AT4 antibody
    • NF-ATc3 antibody
    • NFAC3_HUMAN antibody
    • NFAT4 antibody
    • NFATc3 antibody
    • NFATx antibody
    • Nuclear factor of activated T cells, cytoplasmic 3 antibody
    • nuclear factor of activated T cells, cytoplasmic, calcineurin dependent 3 antibody
    • Nuclear factor of activated T-cells antibody
    • T cell transcription factor NFAT4 antibody
    • T-cell transcription factor NFAT4 antibody
    see all

Images

  • All lanes : Anti-NFAT4/NF-ATc3 antibody (ab93628) at 1 µg/ml

    Lane 1 : Human thymus tissue lysate - total protein (ab30146)
    Lane 2 : Human kidney tissue lysate - total protein (ab30203)
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 5 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 116 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa, 55 kDa, 70 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes


    NFAT4/NF-ATc3 has six isoforms (Swissprot data). Ab93628 is predicted to cross react with isoforms 1-4. This may explain the banding pattern observed here. Isoforms 2 and 3 are 115-kDa. Isoforms 1 and 4 are 116- and 113-kDa respectively. The band around 80-kDa corresponds to the predicted MW of isoform 6. However, ab93628 is not predicted to react with this isoform. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • ab93628 staining NFAT4/NF-ATc3 in Jurkat cells. The cells were fixed with 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions.  The cells were then incubated with the antibody ab93628 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • IHC image of NFAT4/NF-ATc3 staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab93628, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Peterson YK  et al. Frizzled-5: a high affinity receptor for secreted frizzled-related protein-2 activation of nuclear factor of activated T-cells c3 signaling to promote angiogenesis. Angiogenesis 20:615-628 (2017). Read more (PubMed: 28840375) »
  • Fontenot E  et al. A novel monoclonal antibody to secreted frizzled-related protein 2 inhibits tumor growth. Mol Cancer Ther 12:685-95 (2013). WB ; Human . Read more (PubMed: 23604067) »
See all 2 Publications for this product

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