Anti-NFAT5 抗体 (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
製品の概要
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製品名
Anti-NFAT5 antibody
NFAT5 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to NFAT5 -
由来種
Rabbit -
特異性
Detects Nuclear Factor of Activated T-cells 5 (NFAT 5). -
アプリケーション
適用あり: IHC-P, WB, ICC/IF, IPmore details -
種交差性
交差種: Mouse, Human, African green monkey -
免疫原
Synthetic peptide corresponding to Human NFAT5 aa 1439-1455 (C terminal).
Sequence:DLLVSLQNQGNNLTGSF
(Peptide available asab4978) -
特記事項
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
アプリケーション
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.
アプリケーション | Abreviews | 特記事項 |
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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 |
ターゲット情報
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機能
Plays a role in the inducible expression of genes. Regulates hypertonicity-induced cellular accumulation of osmolytes. -
組織特異性
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. -
配列類似性
Contains 1 RHD (Rel-like) domain. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 10725 Human
- Entrez Gene: 54446 Mouse
- Omim: 604708 Human
- SwissProt: O94916 Human
- SwissProt: Q9WV30 Mouse
- Unigene: 371987 Human
- Unigene: 390057 Mouse
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別名
- Glutamine rich protein H65 antibody
- KIAA0827 antibody
- NF AT5 antibody
see all
画像
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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 kDaWestern 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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
プロトコール
データシートおよび資料
参考文献 (20)
ab3446 は 20 報の論文で使用されています。
- 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