NFATC2/NFAT1 Antibody (25A10.D6.D2) - BSA Free

Immunocytochemistry/Immunofluorescence: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - 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 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, 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 at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye.

Immunohistochemistry-Paraffin: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Both normal and cancer biopsies of deparaffinized Human spleen tissue tissues.

Chromatin Immunoprecipitation: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - IP performed using Matrix ChIP assay with 1.0ul/100ul well of NFATc2 mab. Chromatin aliquots from ~1x10^5 cells used per ChIP pull-down. Quantitative PCR data done x4 using 1ul of eluted DNA in 2ul SYBR real-time PCR reactions containing primers to amplify exon-1 or exon-28 of LAMC1. PCR calibration curves generated per primer pair from a series of sheared total genomic DNA. ChIP presented relative to the total amount of input chromatin. Results represent mean +/- SEM for 3 experiments. Representation of rat LAMC1 locus shown above data where boxes represent exons (black = translated regions, white = untranslated regions); zigzag line represents an intron, & straight line represents upstream sequence. Regions amplified by LAMC1 primers are represented by black bars. Data courtesy of the Innovators Program.

Immunocytochemistry/Immunofluorescence: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Analysis of NFATc2 using NFATc2 Monoclonal Antibody (25A10.D6.D2) shows staining in Hela Cells. NFATc2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing NFATc2 at a dilution of 1:20 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunocytochemistry/Immunofluorescence: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Analysis of NFATc2 using NFATc2 Monoclonal Antibody (25A10.D6.D2) shows staining in MCF-7 Cells. NFATc2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing NFATc2 at a dilution of 1:20 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunocytochemistry/Immunofluorescence: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Analysis of NFATc2 using NFATc2 Monoclonal Antibody (25A10.D6.D2) shows staining in U251 Cells. NFATc2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing NFATc2 at a dilution of 1:20 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunohistochemistry-Paraffin: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Cancer biopsies of deparaffinized Human colon carcinoma tissues.

Immunohistochemistry-Paraffin: NFATC2/NFAT1 Antibody (25A10.D6.D2) [NB300-504] - Both normal and cancer biopsies of deparaffinized Human tonsil tissues.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA, GS, ICC/IF, IHC, IP, ChIP
• Clone: 25A10.D6.D2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

NFATC2/NFAT1 Antibody (25A10.D6.D2) - BSA Free Summary

• 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.
• Specificity: Detects nuclear factor of activated T-cells (NFAT). This does not cross react with NFAT2 (NFATc, NFATc1).
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: NFATC2
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1-3 ul
  • Gel Super Shift Assays 1:1 - 1:100
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:1000
  • Immunohistochemistry
  • Immunohistochemistry-Frozen 1:50 - 1:2000
  • Immunohistochemistry-Paraffin 1:50 - 1:2000
  • Immunoprecipitation 1:10 - 1:500
  • Sandwich ELISA 1:100 - 1:2000
  • Western Blot 1 ug/ml
• Application Notes: ChIP usage was reported in scientific literature (PMID: 15347678). In WB: Detects an approx. 140 kDa protein representing phosphorylated NFAT1 in restiing immune cell and approx. 120 kDa protein in stimulated cells that represents a fully dephosphorylated NAFT1. Use in Immunohistochemistry reported in multiple pieces of scientific literature.
• Theoretical MW: 140 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for NFATC2/NFAT1 Antibody (25A10.D6.D2) - BSA Free

• NFAT pre-existing subunit
• NFAT transcription complex, preexisting component
• NFAT1
• NFAT1nuclear factor of activated T-cells, cytoplasmic 2
• NFATC2
• NF-ATc2
• NFATP
• NF-ATp
• NFATPnuclear factor of activated T-cells, preexisting component
• nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 2
• preexisting nuclear factor of activated T-cells 2
• T cell transcription factor NFAT1
• T-cell transcription factor NFAT1

Background

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, NFAT2, NFAT3, and NFAT4. 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NFATC2/NFAT1 Antibody (NB300-504)(13)

Publications using NB300-504

• Wang M, Zhao M, Xu S et al. TRPA1 deficiency attenuates cardiac fibrosis via regulating GRK5/NFAT signaling in diabetic rats Biochemical pharmacology 2023-06-26 [PMID: 37380112]
• Zhu L, Zhou X, Gu M et al. Dapl1 controls NFATc2 activation to regulate CD8+ T cell exhaustion and responses in chronic infection and cancer Nature cell biology 2022-07-01 [PMID: 35773432] (Chemotaxis, Mouse)
• Li HF, Wu YL, Tseng TL, Chao SW et Al. Inhibition of miR-155 potentially protects against lipopolysaccharide-induced acute lung injury through the IRF2BP2-NFAT1 pathway Am J Physiol Cell Physiol 2020-10-14 [PMID: 33052070]
• Song S, Carr SG, McDermott KM et al. STIM2 (Stromal Interaction Molecule 2)-Mediated Increase in Resting Cytosolic Free Ca2+ Concentration Stimulates PASMC Proliferation in Pulmonary Arterial Hypertension Hypertension 2018-01-22 [PMID: 29358461] (Human)
• Younan P, Iampietro M, Nishida A et al. Ebola Virus Binding to Tim-1 on T Lymphocytes Induces a Cytokine Storm. MBio 2017-09-26 [PMID: 28951472] (WB, Mouse)
• Liu J, Han Z, Han Z, He Z Mesenchymal stem cell-conditioned media suppresses inflammation-associated overproliferation of pulmonary artery smooth muscle cells in a rat model of pulmonary hypertension. Exp Ther Med 2016-02-01 [PMID: 26893632] (IF/IHC)
• Cotroneo E, Ashek A, Wang L et al. Iron Homeostasis and Pulmonary Hypertension: Iron Deficiency Leads to Pulmonary Vascular Remodeling in the Rat. Circ. Res. 2015-03-12 [PMID: 25767292] (IF/IHC, WB, Rat)
• Iampietro M, Morissette G, Gravel A, Flamand L. Inhibition of Interleukin-2 Gene Expression by Human Herpesvirus 6B U54 Tegument Protein. J Virol. 2014-08-13 [PMID: 25122797] (WB, Human)
• Landry JW, Banerjee S, Taylor B et al. Chromatin remodeling complex NURF regulates thymocyte maturation. Genes Dev;25(3):275-86. 2011-02-01 [PMID: 21289071]
• Alpini G, Franchitto A, Demorrow S et al. Activation of alpha(1) -adrenergic receptors stimulate the growth of small mouse cholangiocytes via calcium-dependent activation of nuclear factor of activated T cells 2 and specificity protein 1 Hepatology 2011-02-01 [PMID: 21274883] (ICC/IF, IF/IHC, Mouse)
• Barlic J, McDermott DH, Merrell MN et al. Interleukin (IL)-15 and IL-2 reciprocally regulate expression of the chemokine receptor CX3CR1 through selective NFAT1- and NFAT2-dependent mechanisms. J Biol Chem. 2004-11-01 [PMID: 15347678]
• Kornau HC et al. Domain interaction between NMDA receptor subunits the postsynaptic density protein PSD-95. Science 269:1737-40. 1995-01-01 [PMID: 7569905]
• Zhao M, Zheng Z, Xu Y et al. TRPA1 deficiency attenuates cardiac fibrosis via regulating GRK5/NFAT signaling in diabetic rats Research Square 2022-08-25 (ICC/IF, WB, Rat)

Bioinformatics

• Gene Symbol: NFATC2
• Entrez:
  • 4773(Human)
  • 18019(Mouse)
  • 311658(Rat)