GLI-1 Antibody [Unconjugated]

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.

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 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.

• Reactivity: Hu
• Applications: WB, IHC, ChIP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

GLI-1 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human GLI-1
  Met1-Glu234
  Accession # P08151
• Specificity: Detects human GLI-1 in direct ELISAs and Western blots. In Western blots, approximately 5% cross-reactivity with recombinant human GLI-3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: GLI1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GLI-1 Antibody [Unconjugated]

• GLI family zinc finger 1
• GLI1
• GLI-1
• glioma-associated oncogene family zinc finger 1
• glioma-associated oncogene homolog 1 (zinc finger protein)
• Glioma-associated oncogene
• GLIzinc finger protein GLI1
• Oncogene GLI

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GLI-1 Antibody (AF3324)(5)

Publications using AF3324

• M Kurtovi?, N Piteša, N Bartoni?ek, P Ozreti?, V Musani, J ?onkaš, T Petri?, C King, M Sabol RNA-seq and ChIP-seq Identification of Unique and Overlapping Targets of GLI Transcription Factors in Melanoma Cell Lines Cancers, 2022-09-19;14(18):. 2022-09-19 [PMID: 36139698] (ChIP, Human)
• T Fukutani, S Toratani, T Kanda, K Matsui, S Yamasaki, K Sumi, I Ogawa, S Yanamoto Two Cases of Temporomandibular Synovial Chondromatosis Associated with Gli1 Gene Mutation International Journal of Environmental Research and Public Health, 2022-04-13;19(8):. 2022-04-13 [PMID: 35457572] (IHC, Human)
• D Hasegawa, H Ochiai-Shi, S Onodera, T Nakamura, A Saito, T Onda, K Watanabe, K Nishimura, M Ohtaka, M Nakanishi, K Kosaki, A Yamaguchi, T Shibahara, T Azuma Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction PLoS ONE, 2017-10-31;12(10):e0186879. 2017-10-31 [PMID: 29088246] (ICC, Human)
• Pablo Berlanga, Lisandra Muñoz, Marta Piqueras, J. Antoni Sirerol, María Dolors Sánchez-Izquierdo, David Hervás et al. miR-200c and phospho-AKT as prognostic factors and mediators of osteosarcoma progression and lung metastasis Molecular Oncology 2016-08-01 [PMID: 27155790]
• Wang CY, Wei Q, Han I Hedgehog and Notch signaling regulate self-renewal of undifferentiated pleomorphic sarcomas. Cancer Res., 2012-01-09;72(4):1013-22. 2012-01-09 [PMID: 22232736] (IHC, Human)

Bioinformatics

• Gene Symbol: GLI1
• Uniprot:
  • P08151()