Detection of Mouse GLI‑3 by Western Blot. Western blot shows lysates of mouse embryo tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse GLI‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3690) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for GLI‑3 at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of GLI‑3-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 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. GLI‑3/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse GLI‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3690) 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 gli-1 promoter was detected by standard PCR.
GLI‑3 in HeLa Human Cell Line. GLI‑3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Human/Mouse GLI‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3690) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
Human GLI-3 is a 190 kDa transcription factor of the C2H2-type
zinc-finger protein family. GLI-3 appears to play a role in limb and
brain development, probably through sonic hedgehog signaling. GLI-3 is
regulated by PKA-mediated phosphorylation. Unphosphorylated full-length
GLI-3 is a transcriptional activator, while phosphorylated GLI-3 is
cleaved after amino acid 700 to generate an N-terminal 83 kDa
transcriptional repressor. Over the region used as immunogen, human
GLI-3 is 95% identical to the corresponding mouse and canine protein
GLI-Kruppel family member GLI3
Entrez Gene IDs:
2737 (Human); 14634 (Mouse); 140588 (Rat)
ACLS; ACLSPAPAPHS; ADD; Bph; GCPS; GLI family zinc finger 3; GLI3 form of 190 kDa; GLI3 full length protein; GLI3; GLI3-190; GLI3FL; GLI-Kruppel family member GLI3; glioma-associated oncogene family zinc finger 3; Greig cephalopolysyndactyly syndrome; oncogene GLI3; PAP-A; PAPA1; PAPB; PAPBDNA-binding protein; Pdn; PHS; PPDIV; Xt; zinc finger protein GLI3
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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