Immunocytochemistry/Immunofluorescence: NFAT5 Antibody [NB120-3446] - Analysis of NFAT5 using anti-NFAT5 polyclonal antibody (shown in green) in HeLa cells.

Immunohistochemistry-Paraffin: NFAT5 Antibody [NB120-3446] - Normal biopsies of deparaffinized human skeletal muscle tissue.

Western Blot: NFAT5 Antibody [NB120-3446] - 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 probed with a rabbit polyclonal antibody recognizing NFAT5 at a dilution of 1:1000 overnight at 4C 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 using Super Signal West Dura.

Immunocytochemistry/Immunofluorescence: NFAT5 Antibody [NB120-3446] - Analysis of NFAT5 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a NFAT5 polyclonal antibody at a dilution of 1:20 overnight at 4C, 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.

Immunocytochemistry/Immunofluorescence: NFAT5 Antibody [NB120-3446] - 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) or with a NFAT5 polyclonal antibody at a dilution of 1:200 overnight at 4C, 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.

Immunocytochemistry/Immunofluorescence: NFAT5 Antibody [NB120-3446] - 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) or with a NFAT5 polyclonal antibody at a dilution of 1:20 overnight at 4C, 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.

Immunohistochemistry-Paraffin: NFAT5 Antibody [NB120-3446] - Normal biopsies of deparaffinized human brain tissue.

Immunoprecipitation: NFAT5 Antibody [NB120-3446] - Analysis of NFAT5 was performed on U2OS cells. The antigen:antibody complex was formed by incubating 500ug whole cell lysate with 3ug of rabbit polyclonal antibody recognizing NFAT5 overnight on a rocking platform at 4C. The immune-complex was captured on 50ul 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 using Super Signal West Dura.