Key features and details
- Rabbit polyclonal to NFATC4
- Suitable for: WB, ICC/IF, IP
- Reacts with: Mouse, Human
- Isotype: IgG
Storage instructionsShipped 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.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
Concentration information loading...
PurityImmunogen 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.
FunctionPlays 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.
Tissue specificityHighly expressed in placenta, lung, kidney, testis and ovary. Weakly expressed in spleen and thymus. Not expressed in peripheral blood lymphocytes. Detected in hippocampus.
Sequence similaritiesContains 1 IPT/TIG domain.
Contains 1 RHD (Rel-like) domain.
DomainRel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
modificationsPhosphorylated 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'.
Cellular localizationCytoplasm. 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 has been referenced in 3 publications.
- 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