Human GLI-1 Antibody

R&D Systems | Catalog # AF3324

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all GLI-1 Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human GLI-1 Antibody

Detection of Human GLI-1 antibody by Western Blot.

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 antibody by Chromatin Immunoprecipitation.

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. TheBcl-2promoter was detected by standard PCR.
GLI-1 antibody in Human Skin by Immunohistochemistry (IHC-P).

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.

Applications for Human GLI-1 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/106 cells
Sample: PMA and ionomycin treated Jurkat human acute T cell leukemia cell line chromatin, Bcl-2 promoter detected by standard PCR

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human skin

Western Blot

1 µg/mL
Sample: HEK293 human embryonic kidney cell line transfected with human GLI-1

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GLI-1

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.

Long Name

Glioma-Associated Oncogene Homolog 1 [Zinc Finger Protein]

Alternate Names

GLI1

Entrez Gene IDs

2735 (Human); 14632 (Mouse); 140589 (Rat)

Gene Symbol

GLI1

UniProt

P08151

Additional GLI-1 Products

Product Documents for Human GLI-1 Antibody

Certificate of Analysis

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Product Specific Notices for Human GLI-1 Antibody

For research use only

Citations for Human GLI-1 Antibody

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Protocols

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