Oct-4 antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) with 30 ug of chromatin from undifferentiated hESC cells and 7 ul of antibody. ChIP DNA was sequenced on the Illumina HiSeq and11 million sequence tags were mapped to identify Oct-4 binding sites. The image shows binding across a region of chromosome 11. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.
Oct-4 antibody (pAb) tested by Immunofluorescence Mouse embryonic stem cells (mESCs) grown on mouse embryonic fibroblast feeder cells (MEFs) were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were then permeabilized and blocked by incubating with Blocking Solution containing 5% serum/0.1% Triton X-100 in D-PBS for 2 hours at room temperature. Cells were then incubated with Oct-4 antibody (Catalog No. 39811, red) at 1:200 dilution overnight at 4°C, washed with D-PBS, and incubated for 2 hours at room temperature with goat anti-mouse Cy3 secondary antibody at 1:250 dilution. Nuclei were stained with DAPI (blue). Cells were visualized using a Zeiss fluorescent microscope at 20X magnification. Images show that Oct-4 antibody specifically stains mESC colonies and does not stain MEFs. Absence of Oct-4 staining in a subset of cells within the colonies suggests differentiation. Scale bars, 100 μm.
Oct-4 antibody tested by Western blot. Detection of Oct-4 by Western blot. P19 cell nuclear extract (25 ug) probed with Oct-4 antibody (pab) (1:1,000 dilution).
