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Immunology Adaptive Immunity T Cells Non-CD

Anti-NFATC4抗体(ab3447)

  • Datasheet
  • SDS
Submit a review Q&A (3)References (5)

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Western blot - Anti-NFATC4 antibody (ab3447)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)
  • Immunoprecipitation - Anti-NFATC4 antibody (ab3447)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)

Key features and details

  • Rabbit polyclonal to NFATC4
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-NFATC4抗体
    参阅全部 NFATC4 一抗
  • 描述

    兔多克隆抗体to NFATC4
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, ICC/IF, IPmore details
  • 种属反应性

    与反应: Human
  • 免疫原

    Synthetic peptide corresponding to Human NFATC4 aa 887-902.
    Sequence:

    RDLSGFPAPPGEEPPA


    (Peptide available as ab4979)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • WB: MCF7, Jurkat, Raji, HeLa cells; ICC/IF: U251, HeLa, A431 cell lines; IP: MCF7 cells
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped 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.
  • 存储溶液

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Non-CD
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFATS
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFATs

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab3447于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 99 kDa).
ICC/IF
1/100.
IP
Use at an assay dependent concentration.

3 μg

说明
WB
1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 99 kDa).
ICC/IF
1/100.
IP
Use at an assay dependent concentration.

3 μg

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 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.
  • 组织特异性

    Highly expressed in placenta, lung, kidney, testis and ovary. Weakly expressed in spleen and thymus. Not expressed in peripheral blood lymphocytes. Detected in hippocampus.
  • 序列相似性

    Contains 1 IPT/TIG domain.
    Contains 1 RHD (Rel-like) domain.
  • 结构域

    Rel Similarity Domain (RSD) allows DNA-binding and cooperative interactions with AP1 factors.
  • 翻译后修饰

    Phosphorylated 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'.
  • 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.
  • Target information above from: UniProt accession Q14934 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 4776 Human
    • Omim: 602699 Human
    • SwissProt: Q14934 Human
    • Unigene: 77810 Human
    • 别名

      • cytoplasmic 4 antibody
      • NF ATc4 antibody
      • NF-AT3 antibody
      • NF-ATc4 antibody
      • NFAC4_HUMAN antibody
      • NFAT3 antibody
      • NFATc4 antibody
      • Nuclear factor of activated T cells cytoplasmic 4 antibody
      • Nuclear factor of activated T cells cytoplasmic calcineurin dependent 4 antibody
      • Nuclear factor of activated T-cells antibody
      • T cell transcription factor NFAT3 antibody
      • T-cell transcription factor NFAT3 antibody
      see all

    图片

    • Western blot - Anti-NFATC4 antibody (ab3447)
      Western blot - Anti-NFATC4 antibody (ab3447)
      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.

      Secondary
      All lanes : HRP conjugated Goat anti-rabbit at 1/20000 dilution

      Predicted band size: 99 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)
      Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)

      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 - Anti-NFATC4 antibody (ab3447)
      Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)

      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 - Anti-NFATC4 antibody (ab3447)
      Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)

      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 - Anti-NFATC4 antibody (ab3447)
      Immunoprecipitation - Anti-NFATC4 antibody (ab3447)

      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

      Secondary
      All lanes : HRP-conjugated Goat anti-rabbit at 1/20000 dilution
    • Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)
      Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (ab3447)

      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.

    实验方案

    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (5)

    发表研究结果有使用 ab3447?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3447 被引用在 5 文献中.

    • Kohno M  et al. Enhancing calmodulin binding to cardiac ryanodine receptor completely inhibits pressure-overload induced hypertrophic signaling. Commun Biol 3:714 (2020). PubMed: 33244105
    • 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
    • Zhao X  et al. [Expression and significance of COX-2 and its transcription factors NFAT3 and c-Jun in non-small cell lung cancer]. Zhongguo Fei Ai Za Zhi 13:1035-40 (2010). PubMed: 21081043

    客户评价及客户问答

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    1-3 of 3 Abreviews or Q&A

    Question

    BATCH NUMBER 306063 ORDER NUMBER 230325 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE human B cell lymphoma cell line whole cell extract PRIMARY ANTIBODY Abcam anti-NFATc4 was used Recommended dilution 1:10 000, as well as 1:5000 used Antibody was diluted in TBST with 1% BSA, and membrane was incubated at 4 deg celcius overnight Membrane was then washed with TBST thrice, 10 min each time on shaker DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Bands obtained using same sample with Santa Cruz's anti-NFATc4 ANTIBODY STORAGE CONDITIONS Antibody was aliquoted and stored at -30 deg celcius upon arrival SAMPLE PREPARATION Proteins were extracted using RIPA buffer with protease inhibitors. Protein was heated with 4x loading dye at 95 deg celcius for 5 mins, before being loaded. AMOUNT OF PROTEIN LOADED 40ug ELECTROPHORESIS/GEL CONDITIONS 6% SDS-PAGE gel used. 80V was used initially when proteins were migrating through the stacking gel, and increased to 140V thereafter. TRANSFER AND BLOCKING CONDITIONS Transfer to PVDF memebrane was carried out at 4 deg celcius overnight at 30V. Transfer buffer composition [2.9g glycine, 5.8g Tris, 0.37g SDS, pH8.3] Membrane was blocked in 5% BSA for 1 h at room temperature. SECONDARY ANTIBODY Santa Cruz Biotechnology goat-anti-rabbit IgG-HRP used Dilution 1:1000 in TBST with 1% BSA Membrane was incubated for 1 h at room temperature, and subsequently washed thrice in TBST, 10 min per wash with agitation HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? As recommended in answer for previous enquiry CCE1098662 - using a human protein sample and replacing the milk with BSA

    Read More

    Abcam community

    Verified customer

    Asked on Sep 04 2007

    Answer

    Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab3447 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked through the protocols you used and would like to confirm the following items. - can you confirm that the second antibody you used also managed to produce good results with other primary antibodies? In some cases, the problem may be with the secondary antibody not working. - could the no signal you are experiencing be due to poor transfer of protein to membrane? Did you check the transfer efficiency? This can be simply done with a reversible stain such as Ponceau S. I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, I will be more than happy to offer you a replacement or refund. In which case, please confirm your shipping address/purchasing agent information. Also, please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund for you.

    Read More

    Abcam Scientific Support

    回复于 Sep 04 2007

    Question

    What methods for antigen retrieval would you recommend after formalin fixation for immunofluorescence?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2005

    Answer

    Thank you for your enquiry. Unfortunately I do not have details of specific antigen retrieval that should be performed for this antibody. However, may I recommend you start with the mildest form of antigen retrieval in the form of heat mediated antigen retrieval. A time course should be performed to determine the best conditions to present the antigen to the antibody. We have excellent protocols located at the following web link: https://www.abcam.com/index.html?pageconfig=antigenretrieval I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    回复于 Dec 05 2005

    Question

    Has the NFAT3 antibody been tested for reactivity with mouse NFAT3?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 13 2003

    Answer

    According to the datasheet, this antibody has not yet been tested in mouse. We will update the on-line datasheet of this product as soon as get more data.

    Read More

    Asdf Edo

    Abcam Scientific Support

    回复于 Oct 13 2003

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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