Key features and details
- Rabbit polyclonal to NFATC4
- Suitable for: WB, ICC/IF, IP
- Reacts with: Mouse, Human
- Isotype: IgG
参阅全部 NFATC4 一抗
经测试应用适用于: WB, ICC/IF, IPmore details
种属反应性与反应: Mouse, Human
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
存储溶液Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
Concentration information loading...
纯度Immunogen affinity purified
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.
|WB||1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 99 kDa).|
|IP||Use at an assay dependent concentration.
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
- cytoplasmic 4 antibody
- NF ATc4 antibody
- NF-AT3 antibody
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.
All lanes : HRP conjugated Goat anti-rabbit at 1/20000 dilution
Predicted band size: 99 kDa
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.
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 :
Lane 1 : MCF7 cell lysate
Lane 2 : Immunoprecipitate
All lanes : HRP-conjugated Goat anti-rabbit at 1/20000 dilution
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.
ab3447 被引用在 4 文献中.
- Dittmann K et al. New roles for nuclear EGFR in regulating the stability and translation of mRNAs associated with VEGF signaling. PLoS One 12:e0189087 (2017). PubMed: 29253018
- Balakrishnan A et al. Temporal Analysis of Gene Expression in the Murine Schwann Cell Lineage and the Acutely Injured Postnatal Nerve. PLoS One 11:e0153256 (2016). IHC . PubMed: 27058953
- 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 . PubMed: 20530871
- Zhao X et al. [Expression and significance of COX-2 and its transcription factors NFAT3 and c-Jun in non-small cell lung cancer]. Zhongguo Fei Ai Za Zhi 13:1035-40 (2010). PubMed: 21081043