Immunofluorescent analysis of NFAT5 using anti-NFAT5 polyclonal antibody (Pierce Product# PA1-023) (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% Blocker BSA (Pierce Product #37525) for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing NFAT5 (Pierce Product# PA1-023), 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 (Pierce Product# 35552) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Pierce Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rabbit, Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C D(1439) L L V S L Q N Q G N N L T G S F(1455) of human NFAT5.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Gel Shift (GS)||Assay dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||3 µg|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-023 detects nuclear factor of activated T-cells 5 (NFAT5) from human and mouse cells as well as recombinant human protein.
PA1-023 has been successfully used in Western blot, immunofluorescence, immunoprecipitation, gel shift and immunocytochemistry procedures. By Western blot, this antibody detects an ~170 kDa protein representing NFAT5 in HEK293 cells transfected with the human NFAT5 gene. Immunocytochemical staining of NFAT5 in HEK293 cells transfected with the human NFAT5 gene with PA1-023 results in primarily cytoplasmic staining.
The PA1-023 immunogen is a synthetic peptide corresponding to residues C D(1439) L L V S L Q N Q G N N L T G S F(1455) of human NFAT5. This sequence is 94% conserved in the mouse protein. PA1-023 immunizing peptide (Cat. # PEP-123) is available for use in neutralization and control experiments.
The nuclear factor of activated T-cells (NFAT) transcription complex is required for the expression of a group of proteins that collectively regulate the immune response. Four NFAT proteins, encoded on separate genes and expressed as several splice variants, have been described: NFAT1 (also known as NFATp or NFATc2), NFAT2 (NFATc or NFATc1), NFAT3, and NFAT4 (NFATx or NFATc3). These proteins show a low level of sequence similarity with the Dorsal/Rel/NFkB family of transcription factors. Another NFAT-related protein termed NFAT5 differs from isoforms 1-4 in that it lacks many of the Fos/Jun contact sites observed in its predecessors and its subcellular localization is not calcineurin-dependent.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Tumour exosome integrins determine organotropic metastasis.
PA1-023 was used in western blot to analyze organotropic metasistasis and tumour exosome integrins
|Hoshino A,Costa-Silva B,Shen TL,Rodrigues G,Hashimoto A,Tesic Mark M,Molina H,Kohsaka S,Di Giannatale A,Ceder S,Singh S,Williams C,Soplop N,Uryu K,Pharmer L,King T,Bojmar L,Davies AE,Ararso Y,Zhang T,Zhang H,Hernandez J,Weiss JM,Dumont-Cole VD,Kramer K,Wexler LH,Narendran A,Schwartz GK,Healey JH,Sandstrom P,Labori KJ,Kure EH,Grandgenett PM,Hollingsworth MA,de Sousa M,Kaur S,Jain M,Mallya K,Batra SK,Jarnagin WR,Brady MS,Fodstad O,Muller V,Pantel K,Minn AJ,Bissell MJ,Garcia BA,Kang Y,Rajasekhar VK,Ghajar CM,Matei I,Peinado H,Bromberg J,Lyden D||Nature (527:329)||2015|
Calcium/calcineurin synergizes with prostratin to promote NF-¿B dependent activation of latent HIV.
PA1-023 was used in western blot to study NFkB-dependent activation of latent HIV infection and the synergistic role of calcineurin and prostratin
|Chan JK,Bhattacharyya D,Lassen KG,Ruelas D,Greene WC||PloS one (8:null)||2013|
Intrauterine growth restriction modifies the normal gene expression in kidney from rabbit fetuses.
PA1-023 was used in western blot to study the effects on fetal rabbit renal gene expression of intrauterine growth restriction
|Figueroa H,Lozano M,Suazo C,Eixarch E,Illanes SE,Carreño JE,Villanueva S,Hernández-Andrade E,Gratacós E,Irarrazabal CE||Early human development (88:899)||2012|
NFAT5 regulates the canonical Wnt pathway and is required for cardiomyogenic differentiation.
PA1-023 was used in western blot to study the cardiomyogenic differentiation of stem cells and the role played by NFAT5
|Adachi A,Takahashi T,Ogata T,Imoto-Tsubakimoto H,Nakanishi N,Ueyama T,Matsubara H||Biochemical and biophysical research communications (426:317)||2012|
Mediator of DNA damage checkpoint 1 (MDC1) contributes to high NaCl-induced activation of the osmoprotective transcription factor TonEBP/OREBP.
PA1-023 was used in western blot to investigate the role of MDC1 in the activation of osmoprotective transcription factor TonEBP/OREBP by high salt concentration
|Kunin M,Dmitrieva NI,Gallazzini M,Shen RF,Wang G,Burg MB,Ferraris JD||PloS one (5:null)||2010|
Contribution of SHP-1 protein tyrosine phosphatase to osmotic regulation of the transcription factor TonEBP/OREBP.
PA1-023 was used in western blot to identify the phosphatase regulating responses to hypertonicity
|Zhou X,Gallazzini M,Burg MB,Ferraris JD||Proceedings of the National Academy of Sciences of the United States of America (107:7072)||2010|
The transcription factor NFAT5 is required for cyclin expression and cell cycle progression in cells exposed to hypertonic stress.
PA1-023 was used in western blot to investigate the role of NFAT5 in cyclin expression and cell proliferation under hypertonic conditions
|Drews-Elger K,Ortells MC,Rao A,López-Rodriguez C,Aramburu J||PloS one (4:null)||2009|
Integrin alpha6beta4 controls the expression of genes associated with cell motility, invasion, and metastasis, including S100A4/metastasin.
PA1-023 was used in western blot to characterize alpha6beta4 intergrin's regulation targets
|Chen M,Sinha M,Luxon BA,Bresnick AR,O'Connor KL||The Journal of biological chemistry (284:1484)||2009|
Analysis of the transcriptional activity of endogenous NFAT5 in primary cells using transgenic NFAT-luciferase reporter mice.
PA1-023 was used in western blot to study the activation of NFAT5 in primary cells and its response to hypertonicity
|Morancho B,Minguillón J,Molkentin JD,López-Rodríguez C,Aramburu J||BMC molecular biology (9:null)||2008|
Neuropathy target esterase catalyzes osmoprotective renal synthesis of glycerophosphocholine in response to high NaCl.
PA1-023 was used in western blot to demonstrate the effect of high NaCl on neuropathy target esterase and its subsequent effect on the production and accumulation of glycerophosphocholine in mammalian renal cells.
|Gallazzini M,Ferraris JD,Kunin M,Morris RG,Burg MB||Proceedings of the National Academy of Sciences of the United States of America (103:15260)||2006|
Phosphatidylinositol 3-kinase mediates activation of ATM by high NaCl and by ionizing radiation: Role in osmoprotective transcriptional regulation.
PA1-023 was used in western blot to investigate the role of phosphatidylinositol 3-kinase in NaCl- or ionizing radiation-induced ATM activation.
|Irarrazabal CE,Burg MB,Ward SG,Ferraris JD||Proceedings of the National Academy of Sciences of the United States of America (103:8882)||2006|
Expression of osmotic stress-related genes in tissues of normal and hyposmotic rats.
PA1-023 was used in western blot to study the expression of TONEBP in control and hyposmotic rats
|Zhang Z,Ferraris JD,Brooks HL,Brisc I,Burg MB||American journal of physiology. Renal physiology (285:F688)||2003|
Activity of the TonEBP/OREBP transactivation domain varies directly with extracellular NaCl concentration.
PA1-023 was used in western blot to investigate the tonicity dependence of the transactivation domain activity of TonEBP/OREBP C terminus.
|Ferraris JD,Williams CK,Persaud P,Zhang Z,Chen Y,Burg MB||Proceedings of the National Academy of Sciences of the United States of America (99:739)||2002|
Arterial wall stress controls NFAT5 activity in vascular smooth muscle cells.
PA1-023 was used in ChIP assay to study the mechanisms by which arterial wall mechanical stretch leads to activation of vascular smooth muscle cell NFAT5
|Scherer C,Pfisterer L,Wagner AH,Hödebeck M,Cattaruzza M,Hecker M,Korff T||Journal of the American Heart Association (3:null)||2014|
NFAT5 represses canonical Wnt signaling via inhibition of ß-catenin acetylation and participates in regulating intestinal cell differentiation.
PA1-023 was used in immunohistochemistry and western blot to study the role of beta-catenin acetylation in the mechanism by which NFAT5 modulates Wnt signaling and the significance for NFAT5 regulation of intestinal cell differentiation
|Wang Q,Zhou Y,Rychahou P,Liu C,Weiss HL,Evers BM||Cell death and disease (4:null)||2013|
NFAT5 is activated by hypoxia: role in ischemia and reperfusion in the rat kidney.
PA1-023 was used in immunohistochemistry and western blot to study the induction of NFAT5 expression and activity by hypoxia and the significance for renal ischemic injury
|Villanueva S,Suazo C,Santapau D,Pérez F,Quiroz M,Carreño JE,Illanes S,Lavandero S,Michea L,Irarrazabal CE||PloS one (7:null)||2012|