Anti-NFAT2抗体[7A6] - ChIP Grade (ab2796)

概述

  • 产品名称Anti-NFAT2抗体[7A6] - ChIP Grade
    参阅全部 NFAT2 一抗
  • 描述
    小鼠单克隆抗体[7A6] to NFAT2 - ChIP Grade
  • 经测试应用适用于: ICC, ChIP, WB, IHC-FoFr, IP, Flow Cyt, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Human, Non Human Primates
  • 免疫原

    Bacterially expressed fusion protein containing NFAT 2 residues 1-654.

性能

应用

Our Abpromise guarantee covers the use of ab2796 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC 1/250.
ChIP Use at an assay dependent concentration. PubMed: 20375015
WB 1/2000. Detects a band of approximately 105 kDa (predicted molecular weight: 101 kDa).
IHC-FoFr Use at an assay dependent concentration. PubMed: 20530871
EMSA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF 1/10 - 1/100.
IHC-P Use at an assay dependent concentration. PubMed: 16947118

靶标

  • 功能Plays a role in the inducible expression of cytokine genes in T-cells, especially in the induction of the IL-2 or IL-4 gene transcription. Also controls gene expression in embryonic cardiac cells. Could regulate not only the activation and proliferation but also the differentiation and programmed death of T-lymphocytes as well as lymphoid and non-lymphoid cells.
  • 组织特异性Expressed in thymus, peripheral leukocytes as T-cells and spleen. Isoforms A are preferentially expressed in effector T-cells (thymus and peripheral leukocytes) whereas isoforms B and isoforms C are preferentially expressed in naive T-cells (spleen). Isoforms B are expressed in naive T-cells after first antigen exposure and isoforms A are expressed in effector T-cells after second antigen exposure.
  • 序列相似性Contains 1 RHD (Rel-like) domain.
  • 结构域Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
    The N-terminal transactivation domain (TAD-A) binds to and is activated by Cbp/p300. The dephosphorylated form contains two unmasked nuclear localization signals (NLS), which allow translocation of the protein to the nucleus.
    Isoforms C have a C-terminal part with an additional trans-activation domain, TAD-B, which acts as a transcriptional activator. Isoforms B have a shorter C-terminal part without complete TAD-B which acts as a transcriptional repressor.
  • 翻译后修饰Phosphorylated by NFATC-kinase; dephosphorylated by calcineurin.
  • 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
  • 数据库链接
  • 别名
    • cytoplasmic 1 antibody
    • MGC138448 antibody
    • NF ATc antibody
    • NF ATc1 antibody
    • NF-ATc antibody
    • NF-ATc1 antibody
    • NF-ATc1.2 antibody
    • NFAC1_HUMAN antibody
    • NFAT 2 antibody
    • NFAT transcription complex cytosolic component antibody
    • NFATC 1 antibody
    • NFATc antibody
    • NFATc1 antibody
    • Nuclear factor of activated T cells cytoplasmic 1 antibody
    • Nuclear factor of activated T cells cytoplasmic calcineurin dependent 1 antibody
    • Nuclear factor of activated T cells cytosolic component 1 antibody
    • nuclear factor of activated T-cells 'c' antibody
    • Nuclear factor of activated T-cells antibody
    see all

Anti-NFAT2 antibody [7A6] - ChIP Grade 图像

  • Immunocytochemistry/Immunofluorescence analysis of NFAT2 shows staining in 293 cells. NFAT2 (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 incubated without (control) or with ab2796 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NFAT2 shows staining in HeLa cells. NFAT2 (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 incubated without (control) or with ab2796 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NFAT2 shows staining in MCF-7 cells. NFAT2 (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 incubated without (control) or with ab2796 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Ab2796 staining Human normal tonsil tissue. Staining is localised to cytoplasm and nucleus.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 101 kDa


    Exposure time : 10 seconds
  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing NFATc1 ab2796 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human skeletal muscle tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing NFATc1 ab2796 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human spleen tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing NFATc1 ab2796 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Overlay histogram showing Jurkat cells stained with ab2796 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2796, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-NFAT2 antibody [7A6] - ChIP Grade (ab2796)参考文献

This product has been referenced in:
  • So JS  et al. 6-Methoxyflavone inhibits NFAT translocation into the nucleus and suppresses T cell activation. J Immunol 193:2772-83 (2014). Read more (PubMed: 25114106) »
  • Keyes BE  et al. Nfatc1 orchestrates aging in hair follicle stem cells. Proc Natl Acad Sci U S A 110:E4950-9 (2013). IHC-Fr, CHIPseq ; Mouse . Read more (PubMed: 24282298) »

See all 12 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hu pluripotent stem cell derived epicardial like)
Permeabilization Yes - saponin
Specification Hu pluripotent stem cell derived epicardial like
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
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Abcam user community

Verified customer

提交于 May 06 2016

Anti-NFAT2 [7A6] antibody - ChIP Grade, ab2796 binds to the cytoplasmic form within amino acids 1-654.

Thank you for your enquiry.

I can confirm that ab2796 NFAT2 antibody [7A6] antibody is sold asascites fluid. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration st...

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Thank you for your enquiry.

The lot we currently have available is #1312558 (GR73411-4).

I hope this information helps.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Primary Human T cells)
Specification Primary Human T cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 4 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37% Formaldehyde
Detection step Real-time PCR
Positive control Primary Human Regulatory T cells stimulated for 24 hours and analysed for NFAT recruitment to a predicted NFAT binding site
Negative control Primary Human T effector cells stimulated for 24 hours and analysed for NFAT recruitment to the same NFAT binding site. Additionally, an AFM intronic region was also used as an additional negative control.
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Abcam user community

Verified customer

提交于 Jun 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (el4)
Loading amount 100000 cells
Specification el4
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Jun 12 2012

Vielen Dank für diese Nachricht.

Wir freuen uns auf Ihre Rückmeldung - wannimmer Sie soweit sind :-)

Thank you for your inquiry. This antibody is unpurified ascites. The natural color of ascites fluid can vary extensively since it is taken directly from the peritoneal cavity. If "ascites" is stated in the purity section of the datasheet me...

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ab2796 detects NFAT2 from transfected human, rat, mouse, and monkey cells. This antibody does not cross react with NFAT1 as determined by Western Blot. The ab2796 immunogen is bacterially expressed glutathione S-transferase (GST) fusion protein contain...

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Thank you for your enquiry, I was able to find out that with ab2796 we have not looked at endogenously expressed NFAT2, only NFAT2 expressed in the cytosol of transfected cells. Would you be interested in an anti-NFAT1 antibody? Ab2722 detects nucl...

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