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
概述
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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 MouseHumanIHC-P HumanIP HumanWB MouseRatHumanAfrican green monkey -
免疫原
Synthetic peptide corresponding to Human NFAT5 aa 1439-1455 (C terminal).
Sequence:DLLVSLQNQGNNLTGSF
(Peptide available asab4978) -
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
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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...
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纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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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
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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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.
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|
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 |
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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 |
靶标
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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. - Information by UniProt
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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
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别名
- Glutamine rich protein H65 antibody
- KIAA0827 antibody
- NF AT5 antibody
see all
图片
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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.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (20)
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