|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.|
Chromatin Immunoprecipitation (ChIP)
|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. The Bcl-2 promoter was detected by standard PCR.|
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