Human/Mouse Oct-3/4 Antibody

Detection of Human Oct-3/4 by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells and NTera-2 human testicular embryonic carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human/Mouse Oct-3/4 Monoclonal Antibody (Catalog # MAB1759) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Oct-3/4 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Oct‑3/4 in BG01V Human Embryonic Stem Cells. Oct‑3/4 was detected in immersion fixed BG01V human embryonic stem cells using Rat Anti-Human/Mouse Oct‑3/4 Monoclonal Antibody (Catalog # MAB1759) at 10 µg/mL for 3 hours at room temperature. Cells were stained using Rhodamine Red-coupled anti-rat IgG (red) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Oct-3/4 by Immunocytochemistry/Immunofluorescence Pertussis toxin does not affect the maintenance of pluripotency pluripotent stem cell colonies or the ability to form all three embryonic germ layers.(A) Pertussis toxin does not affect the maintenance of pluripotency of hES or hiPS cells as assessed by the expression of the pluripotency markers Nanog, Oct4, Sox2, TDGF1, and ZFP42 by quantitative real-time PCR. (B) Immunocytochemistry of TRA-1-81 (green) and Oct-3/4 (red) also did not reveal differences between hES or hiPS cells treated with pertussis toxin and control treatments in pluripotent stem cell colonies. Nuclei were stained with Hoechst (blue). Scale bar, 200 µm. (C) hES or iPS cells treated with pertussis toxin formed embryoid bodies and differentiated into cell types of the all three embryonic germ layers as assessed by immunocytochemistry for alpha -fetoprotein (a marker of endoderm), alpha -smooth muscle actin (mesoderm) and beta III-tubulin (ectoderm). Scale bar, 100 µm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/19936228), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Oct-3/4 by Western Blot Effect of DFX on spherogenicity of human cancer cell lines and treatment with Nanog siRNA in vitro. (A) After treatment with 0.2% DMSO or 50 μM DFX, a single suspension of HSC-2 cells or OE33 cells was used for the sphere formation assay in a 96-well ultra-low attachment plate. DFX suppressed the spherogenicity of HSC-2 cells and OE33 cells. (B) A single suspension of HSC-2 cells or OE33 cells as described above was used for the spheroid colony assay in a 24-well ultra-low attachment plate. The number of spheres over 50 μm in diameter was counted. The experiments were performed in triplicate, and means ± S.E.M. of each group are shown. DFX significantly suppressed the number of spheres. * p < 0.05. (C) HSC-2 cells were transfected with control or si-Nanog for 48 h, and the expression of stemness markers (Nanog, Sox2, Oct3/4, Klf4, c-Myc) was determined with western blot analysis. beta -actin was used as a loading control. siRNA suppressed the expression of Nanog, Oct3/4, and Klf4. (D) HSC-2 cells were transfected with control or si-Nanog for 48 h, and the sphere formation assay was performed. No differences were found in spherogenicity between the control and si-Nanog cultures. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30717462), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Oct-3/4 by Immunocytochemistry/Immunofluorescence Undifferentiated H7 hESC colonies (on passage 15) 3 days after passaging on hFF feeder cells (a) and after 55 passages on MEF feeder cells (b). H7 cells passaged 11 times on hFF feeder cells expressed Nanog (c and d), Oct4 (e and f), SSEA4 (g and h), and TRA-1-60 (i and j). H7 cells on passage 45 on MEF feeder cells expressed Nanog (k and l), Oct4 (m and n), and SSEA4 (o and p). Scale bars 200 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22315618), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Oct-3/4 by Immunocytochemistry/Immunofluorescence Undifferentiated H7 hESC colonies (on passage 15) 3 days after passaging on hFF feeder cells (a) and after 55 passages on MEF feeder cells (b). H7 cells passaged 11 times on hFF feeder cells expressed Nanog (c and d), Oct4 (e and f), SSEA4 (g and h), and TRA-1-60 (i and j). H7 cells on passage 45 on MEF feeder cells expressed Nanog (k and l), Oct4 (m and n), and SSEA4 (o and p). Scale bars 200 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22315618), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Oct-3/4 by Western Blot The effect of DFX against stemness of miPS cells in vitro and cytotoxicity analysis. (A) miPS cells were treated with the indicated dose of DFX (0, 1, 10, 50, 100 μM) and subjected to western blot analysis with antibodies to stemness markers (Nanog, Sox2, Oct3/4, Klf4, c-Myc) or beta -actin (loading control). Stemness markers were suppressed by DFX at concentrations over 50 μM. (B) miPS cells treated with 50 μM DFX were cultured in suspension for 72 h. DFX treatment of miPS cells suppressed spherogenesis and GFP expression, which indicates suppression of Nanog. (C) Micrographs of the fluorescence-based Live/Dead assay showing live and dead miPS cells following treatment with 0.2% DMSO (control) or 50 μM DFX (magnification ×40). The morphology of miPS cells after treatment with DFX changed from round to spindle shaped. Almost all cells were stained green, which indicates live cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30717462), licensed under a CC-BY license. Not internally tested by R&D Systems.