Human GLI‑1 Antibody

Detection of Human GLI‑1 by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with human GLI-1. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human GLI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3324) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GLI-1 at approximately 165 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of GLI‑1-regulated Genes by Chromatin Immunoprecipitation.

Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 mintes were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. GLI-1/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human GLI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3324) 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. TheBcl-2promoter was detected by standard PCR.

GLI-1 in Human Skin.

GLI-1 was detected in immersion fixed paraffin-embedded sections of human skin using Human GLI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3324 ) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.