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Immunology Adaptive Immunity T Cells Non-CD

Anti-NFAT5抗体(ab3446)

  • Datasheet
  • SDS
Reviews (1)Q&A (1)References (20)

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Western blot - Anti-NFAT5 antibody (ab3446)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
  • Immunoprecipitation - Anti-NFAT5 antibody (ab3446)
  • Immunoprecipitation - Anti-NFAT5 antibody (ab3446)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)

Key features and details

  • Rabbit polyclonal to NFAT5
  • Suitable for: IHC-P, WB, ICC/IF, IP
  • Reacts with: Mouse, Human, African green monkey
  • Isotype: IgG

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概述

  • 产品名称

    Anti-NFAT5抗体
    参阅全部 NFAT5 一抗
  • 描述

    兔多克隆抗体to NFAT5
  • 宿主

    Rabbit
  • 特异性

    Detects Nuclear Factor of Activated T-cells 5 (NFAT 5).
  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    African green monkey
    See all applications and species data
  • 免疫原

    Synthetic peptide corresponding to Human NFAT5 aa 1439-1455 (C terminal).
    Sequence:

    DLLVSLQNQGNNLTGSF


    (Peptide available as ab4978)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators

相关产品

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • NFAT5 peptide (ab4978)
  • Isotype control

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

    • Recombinant Human NFAT5 protein (ab112396)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab3446 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
ICC/IF
Mouse
Human
IHC-P
Human
IP
Human
WB
Mouse
Rat
Human
African green monkey
应用 Ab评论 说明
IHC-P
1/20. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (1)
1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 160 kDa).Can be blocked with NFAT5 peptide (ab4978).
ICC/IF
1/100.
IP
Use at an assay dependent concentration.

3 μg

说明
IHC-P
1/20. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 160 kDa).Can be blocked with NFAT5 peptide (ab4978).
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 genes. Regulates hypertonicity-induced cellular accumulation of osmolytes.
  • 组织特异性

    Highest levels in skeletal muscle, brain, heart and peripheral blood leukocytes. Also expressed in placenta, lung, liver, kidney, pancreas, spleen, thymus, prostate, testis, ovary, small intestine and colon.
  • 序列相似性

    Contains 1 RHD (Rel-like) domain.
  • 细胞定位

    Nucleus.
  • Target information above from: UniProt accession O94916 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: 10725 Human
    • Entrez Gene: 54446 Mouse
    • Omim: 604708 Human
    • SwissProt: O94916 Human
    • SwissProt: Q9WV30 Mouse
    • Unigene: 371987 Human
    • Unigene: 390057 Mouse
    • 别名

      • Glutamine rich protein H65 antibody
      • KIAA0827 antibody
      • NF AT5 antibody
      • NF-AT5 antibody
      • NFAT 5 antibody
      • NFAT L1 antibody
      • NFAT like protein 1 antibody
      • NFAT5 antibody
      • NFAT5_HUMAN antibody
      • NFATL 1 antibody
      • NFATL1 antibody
      • NFATZ antibody
      • Nuclear factor of activated T cells 5 antibody
      • Nuclear factor of activated T cells 5 tonicity responsive antibody
      • Nuclear factor of activated T cells antibody
      • Nuclear factor of activated T-cells 5 antibody
      • OREBP antibody
      • Osmotic response element binding protein antibody
      • T cell transcription factor NFAT 5 antibody
      • T cell transcription factor NFAT5 antibody
      • T-cell transcription factor NFAT5 antibody
      • TonE binding protein antibody
      • TonE-binding protein antibody
      • TonEBP antibody
      • Tonicity responsive enhancer binding protein antibody
      • Tonicity-responsive enhancer-binding protein antibody
      see all

    图片

    • Western blot - Anti-NFAT5 antibody (ab3446)
      Western blot - Anti-NFAT5 antibody (ab3446)
      All lanes : Anti-NFAT5 antibody (ab3446) at 1/1000 dilution

      Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
      Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
      Lane 4 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
      Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 6 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate
      Lane 7 : HeLa (Human epithelial adenocarcinoma cell line) whole cell lysate
      Lane 8 : COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate
      Lane 9 : EL4 (Mouse thymic lymphoma cell line) whole cell lysate
      Lane 10 : C2C12 (Mouse myoblast cell line) whole cell lysate
      Lane 11 : NRK (Rat kidney normal tissue) whole cell lysate

      Lysates/proteins at 25 µg per lane.

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

      Predicted band size: 160 kDa



      Western blot analysis of NFAT5 was performed by loading samples onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were incubated with ab3446 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a secondary antibody for at least one hour. Membranes were washed and chemiluminescent detection performed.

    • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)

      Immunofluorescence analysis of NFAT5 in HeLa (Human epithelial adenocarcinoma cell line) cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3446 at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFAT5 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)

      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 duution with ab3446 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.

    • Immunoprecipitation - Anti-NFAT5 antibody (ab3446)
      Immunoprecipitation - Anti-NFAT5 antibody (ab3446)

      Immunoprecipitation of NFAT5 was performed on U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate (lane 2). The antigen:antibody complex was formed by incubating 500 µg whole cell lysate with 3 µg of ab3446 overnight on a rocking platform at 4°C. The immune-complex was captured on 50 µL Protein A/G Plus 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 1/20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed.

      Lane 1: Only cell lysate.

    • Immunoprecipitation - Anti-NFAT5 antibody (ab3446)
      Immunoprecipitation - Anti-NFAT5 antibody (ab3446)

      Immunoprecipitation of NFAT5 was performed on U2OS cells. The antigen:antibody complex was formed by incubating 500µg whole cell lysate with 3µg of ab3446 overnight on a rocking platform at 4°C. The immune-complex was captured on 50µl Protein A/G Plus 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 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed.

    • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)

      Immunofluorescence analysis of NFAT5 in NIH/3T3 (Mouse embryo fibroblast cell line) cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3446 at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFAT5 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)

      Immunocytochemistry/Immunofluorescence analysis of NFAT5 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 for 15 minutes at room temperature. Cells were probed with ab3446 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)

      Immunohistochemistry was performed on normal biopsies of deparaffinized human brain 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 ab3446 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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFAT5 antibody (ab3446)Image from Tsai TT et al., J Biol Chem. 2006 Sep 1;281(35):25416-24. Epub 2006 Jun 13. Fig 1.; doi: 10.1074/jbc.M601969200; September 1, 2006, The Journal of Biological Chemistry, 281, 25416-25424.
      Immunohistochemical analysis of rat spinal tissue, staining NFAT5 with ab3446. Sections were incubated with primary antibody (1/100) overnight at 4°C before incubating with a biotinylated secondary antibody. Staining was detected using DAB.
    • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)

      Immunofluorescence analysis of NFAT5 in MCF7 (Human breast adenocarcinoma cell line) cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3446 at a dilution of 1:200 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFAT5 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

    • Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunocytochemistry/ Immunofluorescence - Anti-NFAT5 antibody (ab3446)
      Immunofluorescent analysis of NFAT5 usingab3446 (shown in green) in HeLa (Human epithelial adenocarcinoma cell line) whole  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 for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing NFAT5, 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 on a Thermo Scientific ArrayScan at 20X magnification.
       
       

    实验方案

    • Immunoprecipitation protocols
    • Immunohistochemistry 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 (20)

    发表研究结果有使用 ab3446?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab3446 被引用在 20 文献中.

    • Tessier S  et al. Arp2/3 inactivation causes intervertebral disc and cartilage degeneration with dysregulated TonEBP-mediated osmoadaptation. JCI Insight 5:N/A (2020). PubMed: 31961823
    • Maeoka Y  et al. NFAT5 up-regulates expression of the kidney-specific ubiquitin ligase gene Rnf183 under hypertonic conditions in inner-medullary collecting duct cells. J Biol Chem 294:101-115 (2019). PubMed: 30413537
    • Zhai S  et al. Amelioration of Lipopolysaccharide-Induced Nephrotic Proteinuria by NFAT5 Depletion Involves Suppressed NF-?B Activity. Inflammation N/A:N/A (2019). PubMed: 30826989
    • Maeoka Y  et al. Renal medullary tonicity regulates RNF183 expression in the collecting ducts via NFAT5. Biochem Biophys Res Commun 514:436-442 (2019). PubMed: 31053298
    • Arnold C  et al. Genetic ablation of NFAT5/TonEBP in smooth muscle cells impairs flow- and pressure-induced arterial remodeling in mice. FASEB J N/A:fj201801594R (2018). PubMed: 30383452
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-NFAT5 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (JEG3)
    Gel Running Conditions
    Reduced Denaturing (8% gel)
    Loading amount
    30 µg
    Specification
    JEG3
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 20 2017

    Question

    Hello,

    I would like to order antibodies for the following proteins:

    FOXJ2
    HAND1
    NFAT5
    Nkx2-3
    Nkx2-4

    All antibodies will be used for western blotting and should react with rat (heart) tissue.

    Could you please let me know which antibodies you would recommend for these proteins please?

    Thank you very much!

    Best wishes,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 23 2012

    Answer

    Thank you for your inquiry.

    We have a number of antibodies in our catalog that might be of interest to you. I would like to direct you to the search facility on our website (www.abcam.com).

    Please enter the name of the protein into the search box at the top of the home page. A list of products will be generated which can be further refined using the filters on the left hand side of the page. Using these filters will help you to identify products that have been tested in the applications and species you are interested in.

    I have conducted a search for you and found the following antibodies:

    FOXJ2 - 2 antibodies but not tested yet in Rat (suitable for Western blot)

    ab22857, ab49160

    HAND1 - 4 antibodies but not tested yet in Rat (suitable for Western blot)

    ab46822, ab52767, ab86009, ab104320



    NFAT5

    ab3446 - reacts with Rat

    ab105886 - predicted to react with Rat





    Nkx2-3

    ab49162 - predicted to react with Rat



    ab66366- predicted to react with Rat





    Nkx2-4

    ab48331 - predicted to react with Rat



    ab49162 - predicted to react with Rat



    ab66366 - predicted to react with Rat


    If you have specific questions about the products identified during the search, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    回复于 Jan 23 2012

    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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