Anti-NFATC4 antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (AB3447)

Immunofluorescent analysis of NFATC4 using anti-NFATC4 polyclonal antibody ( ab3447) (shown in green) 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% BSA for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing NFATC4 ( ab3447) 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (AB3447)

Immunocytochemistry/Immunofluorescence analysis of NFATC4 in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3447 at a dilution of 1 : 20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (AB3447)

Immunocytochemistry/Immunofluorescence analysis of NFATC4 in A431 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control - right) or with ab3447 at a dilution of 1 : 100 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NFATC4 antibody (AB3447)

Immunocytochemistry/Immunofluorescence analysis of NFATC4 in U251 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control - right) or with ab3447 at a dilution of 1 : 20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. NFATC4 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• IP

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Immunoprecipitation - Anti-NFATC4 antibody (AB3447)

Immunoprecipitation of NFATC4 was performed on MCF7 cells. The antigen : antibody complex was formed by incubating 500μg whole cell lysate with 3μg of rabbit polyclonal antibody recognizing NFATC4 (ab3447) overnight on a rocking platform at 4°C. The immune-complex was captured on 50μl Protein A/G 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 : 20000 for at least one hour. Membranes were washed and chemiluminescent detection performed.

All lanes:

Immunoprecipitation - Anti-NFATC4 antibody (ab3447)

Lane 1:

MCF7 cell lysate

Lane 2:

Immunoprecipitate

Secondary

All lanes:

HRP-conjugated Goat anti-rabbit at 1/20000 dilution

Predicted band size: 95 kDa

false

• WB

Supplier Data

Western blot - Anti-NFATC4 antibody (AB3447)

All lanes:

Western blot - Anti-NFATC4 antibody (ab3447) at 1/1000 dilution

Lane 1:

MCF7 whole cell lysate at 25 µg

Lane 2:

Jurkat whole cell lysate at 25 µg

Lane 3:

Raji whole cell lysate at 25 µg

Lane 4:

Ramos whole cell lysate at 25 µg

Lane 5:

HepG2 whole cell lysate at 25 µg

Lane 6:

U2OS whole cell lysate at 25 µg

Lane 7:

HeLa whole cell lysate at 25 µg

Secondary

All lanes:

HRP conjugated Goat anti-rabbit at 1/20000 dilution

Predicted band size: 95 kDa

false