Mouse Nanog Antibody

Detection of Nanog in D3 Mouse Cell Line by Flow Cytometry.

D3 mouse embryonic stem cell line was stained with Goat Anti-Mouse Nanog Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2729, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).

Nanog in D3 Mouse Cell Line.

Nanog was detected in immersion fixed D3 mouse embryonic stem cell line using Goat Anti-Mouse Nanog Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2729) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Detection of Mouse Nanog by Immunocytochemistry/Immunofluorescence

HP1 beta is highly expressed and diffuse in nuclei of pluripotent cells. a Confocal images of MEFs (top), R1 ESCs (middle) and Rr5 iPSCs (bottom) immunostained for Nanog (green, middle), HP1 beta (red, right) and counterstained with DAPI (blue, left). Asterisks indicate examples of MEFs used as a feeder layer in the culture of the pluripotent cells. b Quantification of the fluorescence intensities of Nanog (green bars) and HP1 beta (red bars) for the three cell types (n ≥ 26). Nanog is used as a marker for pluripotent cells; the fluorescence intensity of the background intensity was subtracted. c Number of HP1 beta foci in the different cell types. Error bars in (b) and (c) represent standard error of the mean. d Confocal images of R1 ESCs immunostained for Nanog (green, middle), HP1 gamma (red, right) and counterstained with DAPI (blue, left). e Confocal images of Rr5 iPSCs immunostained for HP1 gamma (red) and Nanog (inset, green). Asterisks indicate feeder layer MEF cells in (d) and (e). Scale bars for (a–e) = 15 μm. f Time lapse spinning disk confocal images of ESCs expressing the endogenous HP1 beta fused to mCherry induced to differentiate with 1 μM of retinoic acid (RA) for 40 hours (see also Additional file 7 for a video) Image collected and cropped by CiteAb from the following publication (https://genomebiology.biomedcentral.com/articles/10.1186/s13059-015-076…), licensed under a CC-BY license. Not internally tested by R&D Systems.