Western blot shows lysates of HEK293 human embryonic kidney cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-HumanGATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GATA‑5 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
GATA‑5 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human GATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) at 1.7 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 Cells on Coverslips.
HeLa human cervical epithelial carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. GATA‑5/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human GATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) 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 mucin4 promoter was detected by standard PCR.