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. The Bcl-2 promoter 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
GLI-1 is a transcription regulator with 5 conserved tandem C2-H2 zinc
finger domains, joined by conserved histidine/cysteine linkers. It is a
known oncogene that was originally isolated in a human glioblastoma.
GLI-1 acts downstream of Hedgehog signaling and is involved in cell
proliferation and pattern formation during embryonic development.
Within the region used as the immunogen, human GLI-1 shares 81% amino
acid sequence homology with mouse GLI-1.
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but also provides information about sample types, species, and experimental conditions.
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