OCT4 Monoclonal Antibody (9B7) MA1-104

货号：MA1-104

规格：100 µg

价格：4070

产品类型：流式抗体

品牌：Thermo Fisher

抗原：OCT4

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG

克隆号：9B7

荧光染料：Purified

类型:	单抗	同型对照：	IgG
免疫原性：	Full length human recombinant protein of human POU5F1 produced in E.coli.	用法：	3-5 µg/1x10^6 cells(Flow);1.0-2.0 µg/mL(ICC);1.0-2.0 µg/mL(IF);1:500-1:5,000(WB)

Product Specific InformationMA1-104 detects Oct4/Pou5F1 in Human embryonic carcinomic samples, but not in adult samples (e.g. HeLa cells). This antibody did not show reactivity in a mouse embryonic cell line (i.e. F9 embryonic carcinoma cells).MA1-104 has been successfully used in FACS, IF, and WB. By Western blot, this antibody detects a band at 40 kDa in human NCCIT and NTERA-2 embryonal carcinoma cells, but not in negative control cells.Target InformationOct4 is a transcription factor which plays a key role in embryonic development, especially during early embryogenesis. Oct4 is one of the four Yamaka factors, necessary for embryonic stem cell pluripotency. A translocation of this gene with the Ewing's sarcoma gene, t(6;22)(p21;q12), has been linked to tumor formation.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

OCT4 Antibody (MA1-104) in IFImmunofluorescence analysis of Oct4/Pou5F1 was done on 70% confluent log phase NTREA-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Oct4/Pou5F1 (9B7) Mouse Monoclonal Antibody (Product # MA1-104) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification. OCT4 Antibody (MA1-104) in FlowFlow cytometry analysis of Oct4 (red histogram) on human NCCIT cells. Cells were harvested, fixed with 4% formaldehyde, washed with PBS, and incubated with an Oct4 monoclonal antibody (Product # MA1-104) at a 1:100 dilution or PBS alone (blue histogram) for 1 hour on ice. For flow analysis, a 30-minute incubation with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) was performed and 10,000 cells were acquired for each sample. OCT4 Antibody (MA1-104) in IFImmunofluorescent analysis of Oct4 (green) in retinoic acid-treated NCCIT cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 0.3% BSA/TBST (Product # 37525) for 15 minutes at room temperature. Cells were probed with an Oct4 monoclonal antibody (Product # MA1-104) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification. **Note: the absence of Oct4 staining in a subset of the NCCIT cells suggests differentiation. In addition to the nuclear staining of Oct4, the antibody also shows some cytoplasmic staining. OCT4 Antibody (MA1-104) in WBWestern blot analysis of Oct4 was performed by loading 75 µg of the indicated whole cell lysates and 5 µL of the Lane Marker Reducing Sample Buffer (Product # 39000) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST (Product # 37525) for at least 1 hour. The membrane was probed with an Oct4 monoclonal antibody (Product # MA1-104) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075). **Note: the antibody is detecting Oct4 in the human embryonal carcinoma cells, but not in negative control HeLa cells. Unknown bands at 80 kDa were also observed with this antibody.

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