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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] - BSA and Azide free (ab283719)

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Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
  • Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
  • Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24994-68] to NFAT4/NF-ATc3 - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] - BSA and Azide free
    See all NFAT4/NF-ATc3 primary antibodies

  • Description

    Rabbit monoclonal [EPR24994-68] to NFAT4/NF-ATc3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt (Intra),ICC/IF,IHC-P or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Jurkat, HeLa and HEK293T whole cell lysate.

  • General notes

    ab283719 is the carrier-free version of ab283690.

    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
    For more information see here.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283719 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 at an assay dependent concentration. Predicted molecular weight: 116 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 116 kDa.
Application notes
Is unsuitable for Flow Cyt (Intra),ICC/IF,IHC-P or IP.

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

    • Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
      Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
      All lanes : Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] (ab283690) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate
      Lane 2 : NFATC3 (NFAT4) knockout HEK293T (human embryonic kidney epithelial cell) whole cell lysate

      Lysates/proteins at 40 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

      Predicted band size: 116 kDa



      This data was developed using ab283690, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 3% NFDM/TBST

      Lanes 1- 2: Merged signal (red and green). Green - ab283690 observed at 140 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

      ab283690 was shown to react with NFATC3 (NFAT4) in wild-type HEK293T cells in western blot. Loss of signal was observed when NFATC3 (NFAT4) knockout cell line ab267300 (knockout cell lysate ab258073) was used. Wild-type HEK293T and NFATC3 (NFAT4) knockout HEK293T 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. ab283690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1/1000 dilution and 1/20,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

      Lysates were made freshly and used in WB immediately to minimize protein degradation.

       

    • Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
      Western blot - Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
      All lanes : Anti-NFAT4/NF-ATc3 antibody [EPR24994-68] (ab283690) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
      Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 : HEK293T (human embryonic kidney epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 116 kDa
      Observed band size: 140-160 kDa why is the actual band size different from the predicted?



      This data was developed using ab283690, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 3% NFDM/TBST

      The molecular weight observed is consistent with what has been described in the literature (PMID: 31988379).

      Lysates were made freshly and used in WB immediately to minimize protein degradation.

      Exposure time: 3 minutes

    • Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)
      Anti-NFAT4/NF-ATc3 antibody [EPR24994-119] - BSA and Azide free (ab283719)

    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.

    Click here to view the general protocols

    References (0)

    Publishing research using ab283719? Please let us know so that we can cite the reference in this datasheet.

    ab283719 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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