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Immunofluorescent 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.
|Tested species reactivity||Mouse , Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||Full length human recombinant protein of human POU5F1 produced in E.coli.|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg x 10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:500 - 1:5,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-104 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.
Oct4 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.
OTF-3, Oct-3/4, Oct3, Otf-4, Oct4, NF-A3, Otf3g, Oct3/4, Otf-3, Otf3, OTF3, Otf4, OTF4, OCT3, DADB-104B20.2, OCT4, Oct-4, Oct-3, Otf3-rs7
POU domain transcription factor OCT4, POU domain, class 5, transcription factor 1, POU-type homeodomain-containing DNA-binding protein, octamer-binding protein 3, octamer-binding protein 4, octamer-binding transcription factor 3, octamer-binding transcription factor-3, Oct-3, Oct-4, OCT3, OCT4, OTF-3, OTF3, OTF4, POU domain transcription factor OCT4, POU5F1, Homo sapiens POU class 5 homeobox 1 transcript variant 1, DADB-104B20.2, OTF-3, OTF3, OTF4, POU-type homeodomain-containing DNA-binding protein, octamer-binding protein 3, octamer-binding protein 4, octamer-binding transcription factor 3, octamer-binding transcription factor-3