Detection of Human SOX2 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 0.5 µg/mL of Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for SOX2 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of SOX2-regulated Genes by Chromatin Immunoprecipitation. BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. SOX2/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The nanog promoter was detected by standard PCR.
SOX2 in BG01V Human Embryonic Stem Cells. SOX2 was detected in immersion fixed BG01V human embryonic stem cells cultured on irradiated mouse embryonic fibroblasts using Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green, upper panel; Catalog # NL003) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
SOX2 in Mouse Cortical Stem Cells. SOX2 was detected in immersion fixed undifferentiated mouse cortical stem cells using Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) 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). Nestin was also detected in stem cells using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (Catalog # MAB2736) and co-stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
SOX2 in Rat Cortical Stem Cells. SOX2 was detected in immersion fixed undifferentiated rat cortical stem cells using Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) 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). Nestin was also detected using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (Catalog # MAB2736) and stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Detection of Human SOX2 by Simple WesternTM. Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera‑2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SOX2 at approximately 53 kDa (as indicated) using 5 µg/mL of Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.