概述

性能

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应用

Our Abpromise guarantee covers the use of ab3447 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 99 kDa).Can be blocked with NFATC4 peptide (ab4979).

By Western blot, this antibody detects an ~150 kDa protein representing NFATC4 in HEK293 cells transfected with the human NFATC4 gene.

IHC-FoFr Use at an assay dependent concentration. PubMed: 20530871
ICC/IF 1/100.
IHC-P 1/3000.
IP Use at an assay dependent concentration.

3 μg

靶标

  • 功能Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2 and IL-4. Transcriptionally repressed by estrogen receptors; this inhibition is further enhanced by estrogen. Increases the transcriptional activity of PPARG and has a direct role in adipocyte differentiation. May play an important role in myotube differentiation. May play a critical role in cardiac development and hypertrophy. May play a role in deafferentation-induced apoptosis of sensory neurons.
  • 组织特异性Highly expressed in placenta, lung, kidney, testis and ovary. Weakly expressed in spleen and thymus. Not expressed in peripheral blood lymphocytes. Detected in hippocampus.
  • 序列相似性Contains 1 IPT/TIG domain.
    Contains 1 RHD (Rel-like) domain.
  • 结构域Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
  • 翻译后修饰Phosphorylated by NFATC-kinases; dephosphorylated by calcineurin. Phosphorylated on Ser-168 and Ser-170 by MTOR, IRAK1, MAPK7 and MAPK14, on Ser-213 and Ser-217 by MAPK8 and MAPK9, and on Ser-289 and Ser-344 by RPS6KA3. Phosphorylated by GSK3B.
    Ubiquitinated, leading to its degradation by the proteasome and reduced transcriptional activity. Ubiquitination and reduction in transcriptional activity can be further facilitated through GSK3B-dependent phosphorylation. Polyubiquitin linkage is mainly through 'Lys-48'.
  • 细胞定位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
  • 数据库链接
  • 别名
    • cytoplasmic 4 antibody
    • NF ATc4 antibody
    • NF-AT3 antibody
    • NF-ATc4 antibody
    • NFAC4_HUMAN antibody
    • NFAT3 antibody
    • NFATc4 antibody
    • Nuclear factor of activated T cells cytoplasmic 4 antibody
    • Nuclear factor of activated T cells cytoplasmic calcineurin dependent 4 antibody
    • Nuclear factor of activated T-cells antibody
    • T cell transcription factor NFAT3 antibody
    • T-cell transcription factor NFAT3 antibody
    see all

Anti-NFATC4 antibody 图像

  • Immunocytochemistry/Immunofluorescence analysis of NFATC4 in A431 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3447 at a dilution of 1:100 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NFATC4 in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3447 at a dilution of 1:20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NFATC4 in U251 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control - right) or with ab3447 at a dilution of 1:20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunofluorescent analysis of NFATC4 using anti-NFATC4 polyclonal antibody ( ab3447) (shown in green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing NFATC4 ( ab3447) at a dilution of 1:100 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Immunoprecipitation of NFATC4 was performed on MCF7 cells. The antigen:antibody complex was formed by incubating 500µg whole cell lysate with 3µg of rabbit polyclonal antibody recognizing NFATC4 (ab3447) overnight on a rocking platform at 4°C. The immune-complex was captured on 50µl Protein A/G Agarose. Captured immune-complexes were washed and proteins eluted with 5X Reducing Sample Loading Dye . Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody at a dilution of 1:20000 for at least one hour. Membranes were washed and chemiluminescent detection performed.

  • All lanes : Anti-NFATC4 antibody (ab3447) at 1/1000 dilution

    Lane 1 : MCF7 whole cell lysate
    Lane 2 : Jurkat whole cell lysate
    Lane 3 : Raji whole cell lysate
    Lane 4 : Ramos whole cell lysate
    Lane 5 : HepG2 whole cell lysate
    Lane 6 : U2OS whole cell lysate
    Lane 7 : HeLa whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    HRP conjugated Goat anti-rabbit at 1/20000 dilution

    Predicted band size : 99 kDa

Anti-NFATC4 antibody (ab3447)参考文献

This product has been referenced in:
  • Gómez-Sintes R & Lucas JJ NFAT/Fas signaling mediates the neuronal apoptosis and motor side effects of GSK-3 inhibition in a mouse model of lithium therapy. J Clin Invest 120:2432-45 (2010). WB, IHC-FoFr ; Mouse . Read more (PubMed: 20530871) »

See 1 Publication for this product

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Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab3447 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the e...

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Thank you for your enquiry. Unfortunately I do not have details of specific antigen retrieval that should be performed for this antibody. However, may I recommend you start with the mildest form of antigen retrieval in the form of heat mediated ant...

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According to the datasheet, this antibody has not yet been tested in mouse. We will update the on-line datasheet of this product as soon as get more data.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"