Immunofluorescent analysis of NFATc2 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were left untreated (left panel) or treated with 1uM staurosporine (right panel) for 3 hours and probed with a NFATc2 monoclonal antibody (Product # MA1-025), at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan and ToxInsight at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues A(51) I S S P S G L A Y P D D V L D Y G L(69) of mouse NFATc2-A, B and C isoforms.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Gel Shift (GS)||Assay dependent|
|Immunocytochemistry (ICC)||1:100 - 1:1000|
|Immunofluorescence (IF)||1:100 - 1:1000|
|Immunohistochemistry (Frozen) (IHC (F))||1:50-1:200|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 µg/ml|
* 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.
|Western Blot (WB)||See 6 publications below|
|ChIP assay (ChIP)||See 2 publications below|
|Flow Cytometry (Flow)||See 1 publications below|
|Gel Shift (GS)||See 5 publications below|
|Immunoprecipitation (IP)||See 1 publications below|
|Immunocytochemistry (ICC)||See 1 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
MA1-025 detects NFATc2 (NFAT1) from mouse, rat and human tissues. This antibody does not cross react with NFAT2 (NFATc, NFATc1).
MA1-025 has been successfully used in Western blot, ChIP, immunocytochemistry, immunoprecipitation, immunofluorescence, immunohistochemistry (frozen and paraffin), and gel shift procedures. By Western blot, this antibody detects an ~140 kDa protein representing phosphorylated NFAT1 in resting immune cells, and an ~120 kDa protein in stimulated cells that represents fully-dephosphorylated NFAT1.
The MA1-025 immunogen is a synthetic peptide corresponding to residues A(51) I S S P S G L A Y P D D V L D Y G L(69) of mouse NFAT1-A, B and C isoforms. The sequence of this N-terminal peptide is ~70% homologous with human NFAT1.
Antibodies to this protein were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell-based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization, including the selection of the most appropriate fluorescent DyLight conjugated secondary antibody, may have to be performed for your high content assay.
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.
Nuclear factor of activated T-cells (NFAT)C2 inhibits Notch receptor signaling in osteoblasts.
MA1-025 was used in western blot to study the ability of NFATc2 to inhibit osteoblast Notch signaling
|Zanotti S,Smerdel-Ramoya A,Canalis E||The Journal of biological chemistry (288:624)||2013|
Calcineurin/NFAT signalling inhibits myeloid haematopoiesis.
MA1-025 was used in western blot to study the role of calcineurin/NFAT signaling in inhibiting myeloid haematopoiesis
|Fric J,Lim CX,Koh EG,Hofmann B,Chen J,Tay HS,Mohammad Isa SA,Mortellaro A,Ruedl C,Ricciardi-Castagnoli P||EMBO molecular medicine (4:269)||2012|
Hypertrophy and transcriptional regulation induced in myogenic cell line L6-C5 by an increase of extracellular calcium.
MA1-025 was used in western blot to investigate the effect of calcium concentration on cell growth and gene expresion in myogenic cell line L6-C5
|De Arcangelis V,Coletti D,Canato M,Molinaro M,Adamo S,Reggiani C,Naro F||Journal of cellular physiology (202:787)||2005|
|Mouse||1:1,000||TRPC3 channels confer cellular memory of recent neuromuscular activity.||Rosenberg P,Hawkins A,Stiber J,Shelton JM,Hutcheson K,Bassel-Duby R,Shin DM,Yan Z,Williams RS||Proceedings of the National Academy of Sciences of the United States of America (101:9387)||2004|
Identification and characterization of a novel nuclear factor of activated T-cells-1 isoform expressed in mouse brain.
MA1-025 was used in western blot to show that NFAT1-D is expressed in the mouse brain
|Plyte S,Boncristiano M,Fattori E,Galvagni F,Paccani SR,Majolini MB,Oliviero S,Ciliberto G,Telford JL,Baldari CT||The Journal of biological chemistry (276:14350)||2001|
The calcineurin-NFAT pathway and muscle fiber-type gene expression.
MA1-025 was used in western blot to investigate the role of calcineurin-NFAT pathway in the regulation of muscle fiber-type gene expression
|Swoap SJ,Hunter RB,Stevenson EJ,Felton HM,Kansagra NV,Lang JM,Esser KA,Kandarian SC||American journal of physiology. Cell physiology (279:C915)||2000|
Lipocalin 2, the TNF-like receptor TWEAKR and its ligand TWEAK act downstream of NFAT1 to regulate breast cancer cell invasion.
MA1-025 was used in ChIP assay to study the regulation of breast cancer invasiveness by NFAT1 and the role played by lipocalin-2, TWEAKR and TWEAK
|Gaudineau B,Fougère M,Guaddachi F,Lemoine F,de la Grange P,Jauliac S||Journal of cell science (125:4475)||2012|
FOXP3 inhibits activation-induced NFAT2 expression in T cells thereby limiting effector cytokine expression.
MA1-025 was used in chromatin immunoprecipitation to study the effect of FOXP3 on NFAT-mediated inflammatory gene expression.
|Torgerson TR,Genin A,Chen C,Zhang M,Zhou B,Añover-Sombke S,Frank MB,Dozmorov I,Ocheltree E,Kulmala P,Centola M,Ochs HD,Wells AD,Cron RQ||Journal of immunology (Baltimore, Md. : 1950) (183:907)||2009|
Cutting edge: Regulatory T cells selectively attenuate, not terminate, T cell signaling by disrupting NF-¿B nuclear accumulation in CD4 T cells.
MA1-025 was used in flow cytometry to investigate the mechanism of the suppression of CD4 T cell signaling by regulatory T cells
|Huang YH,Sojka DK,Fowell DJ||Journal of immunology (Baltimore, Md. : 1950) (188:947)||2012|
NFATc1 regulation of TRAIL expression in human intestinal cells.
MA1-025 was used in EMSA to investigate the regulatory effect of NFATc1 on TRAIL in human intestinal cells
|Wang Q,Zhou Y,Weiss HL,Chow CW,Evers BM||PloS one (6:null)||2011|
Calcium-dependent activation of interleukin-21 gene expression in T cells.
MA1-025 was used in EMSA to demonstrate the induction of the IL-21 in preactivated T lymphocytes by a calcium signal.
|Kim HP,Korn LL,Gamero AM,Leonard WJ||The Journal of biological chemistry (280:25291)||2005|
Regulation of the murine Nfatc1 gene by NFATc2.
MA1-025 was used in EMSA to investigate the regulatory effect of NFATc2 on the murine Nfatc1 gene.
|Zhou B,Cron RQ,Wu B,Genin A,Wang Z,Liu S,Robson P,Baldwin HS||The Journal of biological chemistry (277:10704)||2002|
A T cell-specific enhancer of the human CD40 ligand gene.
MA1-025 was used in EMSA assay to study the transcriptional regulation of human CD40 ligand gene
|Schubert LA,Cron RQ,Cleary AM,Brunner M,Song A,Lu LS,Jullien P,Krensky AM,Lewis DB||The Journal of biological chemistry (277:7386)||2002|
Double-stranded RNA regulates IL-4 expression.
MA1-025 was used in EMSA to investigate the role of the low concentrations of dsRNA towards the IL-4 expression.
|Kehoe KE,Brown MA,Imani F||Journal of immunology (Baltimore, Md. : 1950) (167:2496)||2001|
Retinoic acid-induced CCR9 expression requires transient TCR stimulation and cooperativity between NFATc2 and the retinoic acid receptor/retinoid X receptor complex.
MA1-025 was used in immunoprecipitation to investigate the cooperativity between NFATc2 and the RAR/RXR and its effect on CCR9 expression
|Ohoka Y,Yokota A,Takeuchi H,Maeda N,Iwata M||Journal of immunology (Baltimore, Md. : 1950) (186:733)||2011|
Pivotal Advance: Nonfunctional lung effectors exhibit decreased calcium mobilization associated with reduced expression of ORAI1.
MA1-025 was used in immunocytochemistry to identify how the function of lung effector cells is inactivated
|Arimilli S,Sharma SK,Yammani R,Reid SD,Parks GD,Alexander-Miller MA||Journal of leukocyte biology (87:977)||2010|
Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway.
MA1-025 was used in immunohistochemistry to demonstrate that nuclear accumulation of NFAT4 is regulated by JNK
|Chow CW,Rincón M,Cavanagh J,Dickens M,Davis RJ||Science (New York, N.Y.) (278:1638)||1997|