GLI-2 Antibody

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‑2/DNA complexes were immunoprecipitated using 5 μg Sheep Anti-Human GLI‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3526) or control antibody (Catalog # 5‑001‑A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). 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.

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

GLI-2 Antibody Summary

• Immunogen: E. coli-derived recombinant human GLI-2
  Ala2-Glu91
  Accession # BAA25668
• Specificity: Detects human GLI-2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 25% cross-reactivity with recombinant mouse GLI-2 and 5% cross-reactivity with recombinant human (rh) GLI-1 and rhGLI-3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: GLI2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 0.1 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-2 Antibody

• GLI family zinc finger 2
• GLI2
• GLI-2
• GLI-Kruppel family member GLI2
• glioma-associated oncogene family zinc finger 2
• HPE9tax helper protein 1
• oncogene GLI2
• tax helper protein 2
• Tax helper protein
• tax-responsive element-2 holding protein
• tax-responsive element-25-bp sequence binding protein
• THP
• THP1
• THP2
• zinc finger protein GLI2

Background

GLI-2 is a 166 kDa biopotential transcription regulator of the hedgehog signaling pathway. It contains 5 conserved tandem C2H2 zinc finger domains that are flanked by a repression domain at the N-terminal region and an activation domain in the C-terminal region. At least four isoforms that differ in their N- or C-terminal regions have been described. Transcriptional activity of GLI-2 is regulated by proteolytic processing and degradation. Within the region used as immunogen, human and mouse GLI-2 share 88% amino acid sequence homology.

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-2 Antibody (AF3526)(7)

Publications using AF3526

• 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)
• Fan Yang, Jezabel Rodriguez-Blanco, Jun Long, Marzena Swiderska-Syn, Daniel T. Wynn, Bin Li et al. A Druggable UHRF1/DNMT1/GLI Complex Regulates Sonic Hedgehog–Dependent Tumor Growth Molecular Cancer Research 2022-11-03 [PMID: 35925047]
• R Taylor, J Long, JW Yoon, R Childs, KB Sylvesters, ML Nielsen, KF Leong, S Iannaccone, DO Walterhous, DJ Robbins, P Iannaccone Regulation of GLI1 by cis DNA elements and epigenetic marks DNA Repair (Amst.), 2019-05-02;79(0):10-21. 2019-05-02 [PMID: 31085420] (Proximity Ligation Assay, Human)
• Raul German Spallanzani, David Zemmour, Tianli Xiao, Teshika Jayewickreme, Chaoran Li, Paul J. Bryce et al. Distinct immunocyte-promoting and adipocyte-generating stromal components coordinate adipose tissue immune and metabolic tenors Science Immunology 2019-05-24 [PMID: 31053654]
• Xin Zheng, Han Han, Guang‐Ping Liu, Yan‐Xiu Ma, Ruo‐Lang Pan, Ling‐Jie Sang et al. LncRNA wires up Hippo and Hedgehog signaling to reprogramme glucose metabolism The EMBO Journal 2017-11-15 [PMID: 28963395]
• Santini R, Pietrobono S, Pandolfi S, Montagnani V, D'Amico M, Penachioni J, Vinci M, Borgognoni L, Stecca B SOX2 regulates self-renewal and tumorigenicity of human melanoma-initiating cells. Oncogene, 2014-03-31;33(38):4697-708. 2014-03-31 [PMID: 24681955] (ChIP, Human)
• Elsawa SF, Almada LL, Ziesmer SC, Novak AJ, Witzig TE, Ansell SM, Fernandez-Zapico ME GLI2 Transcription Factor Mediates Cytokine Cross-talk in the Tumor Microenvironment. J. Biol. Chem., 2011-03-18;286(24):21524-34. 2011-03-18 [PMID: 21454528] (ChIP, Human)

Bioinformatics

• Gene Symbol: GLI2
• Uniprot:
  • BAA25668()