Overview

  • Product name
    Anti-NFAT5 antibody - ChIP Grade
    See all NFAT5 primary antibodies
  • Description
    Rabbit polyclonal to NFAT5 - ChIP Grade
  • Specificity
    Detects Nuclear Factor of Activated T-cells 5 (NFAT 5).
  • Tested applications
    Suitable for: ICC, WB, ChIP, ICC/IF, IP, IHC-P, Immunomicroscopymore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

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

    DLLVSLQNQGNNLTGSF


    (Peptide available as ab4978)

Properties

Applications

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.

Application Abreviews Notes
ICC 1/1000.
WB 1/1000. Detects a band of approximately 170 kDa (predicted molecular weight: 160 kDa).Can be blocked with NFAT5 peptide (ab4978).
ChIP Use at an assay dependent concentration. PubMed: 17513763
ICC/IF 1/100.
IP Use at an assay dependent concentration.

3 μg

IHC-P 1/100. PubMed: 16772300
EMSA Use at an assay dependent concentration.
Immunomicroscopy 1/200. PubMed: 16772300

Target

  • Function
    Plays a role in the inducible expression of genes. Regulates hypertonicity-induced cellular accumulation of osmolytes.
  • Tissue specificity
    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.
  • Sequence similarities
    Contains 1 RHD (Rel-like) domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • 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 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) 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 incubated with ab3446 at a dilution of 1:20 (left) or without primary antibody (negative control - right) 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.



  • Predicted band size : 160 kDa

    Western blot analysis of NFAT5 was performed by loading 25ug of various whole cell lysates 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 at a dilution of 1:1000 overnight at 4°C on a rocking platform. 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 analysis of NFAT5 in HeLa 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) 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 incubated with ab3446 at a dilution of 1:20 (left) or without primary antibody (negative control - right) 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.



  • Predicted band size : 160 kDa

    Western blot of Human NFAT5 from transfected BHK cell lysate with ab3446.

  • Immunocytochemistry/Immunofluorescence analysis of NFAT5 in NIH-3T3 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.

  • 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 analysis of NFAT5 in MCF-7 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.

References

This product has been referenced in:
  • Li C  et al. NFAT5 participates in seawater inhalation-induced acute lung injury via modulation of NF-?B activity. Mol Med Rep 14:5033-5040 (2016). WB, IHC-P ; Rat . Read more (PubMed: 27779669) »
  • Kitterer D  et al. Activation of nuclear factor of activated T cells 5 in the peritoneal membrane of uremic patients. Am J Physiol Renal Physiol 308:F1247-58 (2015). Read more (PubMed: 25834072) »

See all 11 Publications for this product

Customer reviews and Q&As

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
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