Anti-NFATC4抗体
参阅全部 NFATC4 一抗
兔多克隆抗体to NFATC4
Rabbit
适用于: WB, ICC/IF, IPmore details
与反应: Human
Synthetic peptide corresponding to Human NFATC4 aa 887-902.
Sequence:
RDLSGFPAPPGEEPPA
(Peptide available as ab4979)
WB: MCF7, Jurkat, Raji, HeLa cells; ICC/IF: U251, HeLa, A431 cell lines; IP: MCF7 cells
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Liquid
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.
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab3447于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | 1/1000. Detects a band of approximately 95 kDa (predicted molecular weight: 99 kDa). | |
| ICC/IF | 1/100. | |
| IP | Use at an assay dependent concentration. 3 μg |
Entrez Gene: 4776 Human
Omim: 602699 Human
SwissProt: Q14934 Human
Unigene: 77810 Human
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Western blot - Anti-NFATC4 antibody (ab3447)
All lanes : Anti-NFATC4 antibody (ab3447) at 1/1000 dilution
Lane 1 : MCF7 whole cell lysate
Lane 2 : Jurkat whole cell lysate
Lane 3 : Raji whole cell lysate
Lane 4 : Ramos whole cell lysate
Lane 5 : HepG2 whole cell lysate
Lane 6 : U2OS whole cell lysate
Lane 7 : HeLa whole cell lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : HRP conjugated Goat anti-rabbit at 1/20000 dilution
Predicted band size: 99 kDa
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.
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.
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.
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 :
Lane 1 : MCF7 cell lysate
Lane 2 : Immunoprecipitate
Secondary
All lanes : HRP-conjugated Goat anti-rabbit at 1/20000 dilution
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.
