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    nfat1-antibody-epr24658-43-bsa-and-azide-free-ab283720.pdf

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Immunology Adaptive Immunity T Cells Non-CD
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RecombinantRabMAb

Recombinant Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)

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  • Certificate of Compliance
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Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
  • Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
  • Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
  • Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
  • Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)

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 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

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Primary
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Anti-NFAT1 (phospho S54) antibody (ab200819)
Knockout
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Human NFATC2 knockout Raji cell line (ab280906)
Primary
Product image
Anti-NFAT1 antibody [EPR24658-43] (ab283691)

View more associated products

Overview

  • Product name

    Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free
    See all NFAT1 primary antibodies
  • Description

    Rabbit monoclonal [EPR24658-43] to NFAT1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WBmore details
    Unsuitable for: 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: Ramos, Raji, and Daudi whole cell lysates Flow cyt-intra: Ramos and Jurkat cells.
  • General notes

    ab283720 is the carrier-free version of ab283691.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    pH: 7.20
    Constituent: 100% PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR24658-43
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Non-CD
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFATS
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFATs
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Anti-NFAT1 antibody [EPR24658-43] (ab283691)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283720 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 100 kDa.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 100 kDa.
Application notes
Is unsuitable for 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, 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
    modifications

    In 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.
  • Target information above from: UniProt accession Q13469 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4773 Human
    • Omim: 600490 Human
    • SwissProt: Q13469 Human
    • Unigene: 713650 Human
    • Alternative names

      • AI607462 antibody
      • cytoplasmic 2 antibody
      • KIAA0611 antibody
      • NF ATc2 antibody
      • NF ATp antibody
      • NF-ATc2 antibody
      • NF-ATp antibody
      • NFAC2_HUMAN antibody
      • NFAT 1 antibody
      • NFAT pre existing subunit antibody
      • NFAT pre-existing subunit antibody
      • NFAT transcription complex, preexisting component antibody
      • NFAT1 antibody
      • NFAT1-D antibody
      • NFATc2 antibody
      • NFATp antibody
      • Nuclear factor of activated T cells cytoplasmic 2 antibody
      • Nuclear factor of activated T cells cytoplasmic calcineurin dependent 2 antibody
      • Nuclear factor of activated T cells pre-existing component antibody
      • Nuclear factor of activated T cells, preexisting component antibody
      • Nuclear factor of activated T-cells antibody
      • Preexisting nuclear factor of activated T cells 2 antibody
      • T cell transcription factor NFAT 1 antibody
      • T-cell transcription factor NFAT1 antibody
      see all

    Images

    • Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      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 kDa



      This data was developed using ab283691, the same antibody clone in a different buffer formulation.

      False 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.

    • Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      Western blot - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      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 kDa



      This data was developed using ab283691, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: This 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

    • Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)

      This data was developed using ab283691, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of 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 compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: Hela (PMID:21078663).
    • Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      Flow Cytometry (Intracellular) - Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)

      This data was developed using ab283691, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of 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) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: LNCaP.
    • Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)
      Anti-NFAT1 antibody [EPR24658-43] - BSA and Azide free (ab283720)

    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

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab283720? Please let us know so that we can cite the reference in this datasheet.

    ab283720 has not yet been referenced specifically in any publications.

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