Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Drosophila, Frog - Xenopus (African Clawed Frog), Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/Immunofluorescence, Immunocytochemistry, Immunoprecipitation, Chromatin Immunoprecipitation, IF/ICC

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all GLI-2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse GLI-2
Met1-Glu416
Accession # NP_001074594

Specificity

Detects mouse GLI-2 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse GLI‑2 Antibody

GLI-2 antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

GLI‑2 in Mouse Embryo.

GLI-2 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using Goat Anti-Mouse GLI-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3635) at 15 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). Lower panel shows a lack of labeling when primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. Specific staining was localized to developing muscle. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of GLI-2 by Western Blot

Detection of GLI-2 by Western Blot

The PY-type nuclear localization signal (PY-NLS) is required for Gli activation.(A) Western blot of cell lysates from the indicated cell lines indicated that Myc-Gli2WT and Myc-Gli2mPY were expressed at comparable levels that were slightly higher than that of endogenous Gli2. (B–C) Normalized mRNA levels of endogenous Gli1 (B) or Patch1 (C) measured by quantitative reverse transcription PCR (RT-qPCR) in control (green fluorescent protein [GFP] short hairpin RNA [shRNA]) or Gli2-depeleted NIH3T3 cells with or without lentiviral infection of the indicated Gli2 constructs. (D) Fractionation of Myc-Gli2WT and Myc-Gli2mPY from the indicated cell lines treated with or without SAG. Quantification of protein level is shown in the bottom panel. (E–F) Immunostaining (E) and quantification (F) of ciliary-localized Myc-Gli2WT or Myc-Gli2mPY in NIH3T3mGli2-shRNA cells treated with or without sonic hedgehog (Shh). N = 50 cells were examined for each Gli construct. Data are means ± SD from 2 independent experiments. **P < 0.01. (G) Gli-luciferase assay was performed in NIH3T3 cells transfected with the indicated constructs. Data are means ± SD from 2 independent experiments. *P < 0.05, NS: not significant. The underlying data for this figure can be found in S1 Data. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28777795), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GLI-2 by Western Blot

Detection of GLI-2 by Western Blot

The PY-type nuclear localization signal (PY-NLS) is required for Gli activation.(A) Western blot of cell lysates from the indicated cell lines indicated that Myc-Gli2WT and Myc-Gli2mPY were expressed at comparable levels that were slightly higher than that of endogenous Gli2. (B–C) Normalized mRNA levels of endogenous Gli1 (B) or Patch1 (C) measured by quantitative reverse transcription PCR (RT-qPCR) in control (green fluorescent protein [GFP] short hairpin RNA [shRNA]) or Gli2-depeleted NIH3T3 cells with or without lentiviral infection of the indicated Gli2 constructs. (D) Fractionation of Myc-Gli2WT and Myc-Gli2mPY from the indicated cell lines treated with or without SAG. Quantification of protein level is shown in the bottom panel. (E–F) Immunostaining (E) and quantification (F) of ciliary-localized Myc-Gli2WT or Myc-Gli2mPY in NIH3T3mGli2-shRNA cells treated with or without sonic hedgehog (Shh). N = 50 cells were examined for each Gli construct. Data are means ± SD from 2 independent experiments. **P < 0.01. (G) Gli-luciferase assay was performed in NIH3T3 cells transfected with the indicated constructs. Data are means ± SD from 2 independent experiments. *P < 0.05, NS: not significant. The underlying data for this figure can be found in S1 Data. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28777795), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GLI-2 by Western Blot

Detection of Mouse GLI-2 by Western Blot

HPP-9 is a long-acting Hedgehog pathway inhibitor.a SHH-GFP cells were treated for 25 h with DMSO, 1 μM HPI-1, or 1 μM HPP-9, before the medium was changed to various dilutions of ShhN-conditioned medium for 27 h. Nuclear GFP levels were quantified using fluorescence microscopy. Representative curves of three independent experiments are shown, with 9–16 images analyzed per condition. b GLI1 and GLI2 levels of cells pre-incubated with DMSO or 1 μM HPP-9 were determined. Representative blots of three independent experiments are shown. c, d The effect of 1 μM HPP-9 or HPI-1 (pre-)incubation on GLI2 ciliary trafficking was assessed through fluorescence microscopy. Representative images for GLI2 trafficking are shown in (c), and all data is quantified in (d). N independent experiments as indicated in the bars, n = 300–500 cilia analyzed per condition. Mean ± SD is plotted, two-way ANOVA, p as indicated. Scalebar 2 μm. e SHH-GFP cells were incubated with the indicated compounds for 25 h, before the medium was changed to ShhN-conditioned medium with or without competitors for 27 h. Nuclear BRD protein levels were determined using high-content fluorescence microscopy. Data shown is from N independent experiments as indicated, with n = 9–16 images analyzed per condition in each experiment. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GLI-2 by Western Blot

Detection of Mouse GLI-2 by Western Blot

Biological evaluation of HPP-9 and its inactive analog.a Chemical structures of HPP-9 and the methylated analog inact-HPP-9. b Representative micrographs showing the dose-dependent inhibition of an Shh-driven GFP reporter by HPP-9, resulting in the dose-response curves shown in (c). Scalebar 30 μm. b, c Curves and images are representative of N independent experiments (as indicated in the table below the graph), with n = 9 or 18 images analyzed per experiment. d, e NIH-3T3 cells were incubated with increasing concentrations of HPP-9 or HPI-1 in the presence of ShhN and lysates were probed for GLI1, GLI2, and GLI3. d shows a representative immunoblot illustrating that both compounds inhibit GLI1 and GLI2 but have no effect on GLI3 processing. e Quantification (mean ± SEM) of GLI1 (N = 3) and GLI2 full-length levels (N = 2) of N independent experiments. f–h qPCR for Ptch1 (f), Gli1 (g), and Gli2 (h) shows that HPP-9 reduces the expression of Hh pathway target genes, while also decreasing basal Gli2 transcript levels without affecting the fold-induction upon pathway stimulation (−: no ShhN, +: with ShhN). Data shown is the mean (f) or mean ± SD (g, h) for N independent experiments as indicated in the bar. f, g One-way ANOVA with Dunnett’s test, p as indicated in the graph, compared to DMSO + ShhN. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GLI-2 by Western Blot

Detection of Mouse GLI-2 by Western Blot

Biological evaluation of HPP-9 and its inactive analog.a Chemical structures of HPP-9 and the methylated analog inact-HPP-9. b Representative micrographs showing the dose-dependent inhibition of an Shh-driven GFP reporter by HPP-9, resulting in the dose-response curves shown in (c). Scalebar 30 μm. b, c Curves and images are representative of N independent experiments (as indicated in the table below the graph), with n = 9 or 18 images analyzed per experiment. d, e NIH-3T3 cells were incubated with increasing concentrations of HPP-9 or HPI-1 in the presence of ShhN and lysates were probed for GLI1, GLI2, and GLI3. d shows a representative immunoblot illustrating that both compounds inhibit GLI1 and GLI2 but have no effect on GLI3 processing. e Quantification (mean ± SEM) of GLI1 (N = 3) and GLI2 full-length levels (N = 2) of N independent experiments. f–h qPCR for Ptch1 (f), Gli1 (g), and Gli2 (h) shows that HPP-9 reduces the expression of Hh pathway target genes, while also decreasing basal Gli2 transcript levels without affecting the fold-induction upon pathway stimulation (−: no ShhN, +: with ShhN). Data shown is the mean (f) or mean ± SD (g, h) for N independent experiments as indicated in the bar. f, g One-way ANOVA with Dunnett’s test, p as indicated in the graph, compared to DMSO + ShhN. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse GLI-2 by Western Blot

Detection of Mouse GLI-2 by Western Blot

HPP-9 is a long-acting Hedgehog pathway inhibitor.a SHH-GFP cells were treated for 25 h with DMSO, 1 μM HPI-1, or 1 μM HPP-9, before the medium was changed to various dilutions of ShhN-conditioned medium for 27 h. Nuclear GFP levels were quantified using fluorescence microscopy. Representative curves of three independent experiments are shown, with 9–16 images analyzed per condition. b GLI1 and GLI2 levels of cells pre-incubated with DMSO or 1 μM HPP-9 were determined. Representative blots of three independent experiments are shown. c, d The effect of 1 μM HPP-9 or HPI-1 (pre-)incubation on GLI2 ciliary trafficking was assessed through fluorescence microscopy. Representative images for GLI2 trafficking are shown in (c), and all data is quantified in (d). N independent experiments as indicated in the bars, n = 300–500 cilia analyzed per condition. Mean ± SD is plotted, two-way ANOVA, p as indicated. Scalebar 2 μm. e SHH-GFP cells were incubated with the indicated compounds for 25 h, before the medium was changed to ShhN-conditioned medium with or without competitors for 27 h. Nuclear BRD protein levels were determined using high-content fluorescence microscopy. Data shown is from N independent experiments as indicated, with n = 9–16 images analyzed per condition in each experiment. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse GLI‑2 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of mouse embryo (15 d.p.c.)

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse GLI‑2

Reviewed Applications

Read 5 reviews rated 3.4 using AF3635 in the following applications:

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-2

Mouse GLI-2 is a 185 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 two additional isoforms that have amino acid substitutions/truncations in the region corresponding to amino acid residues 192‑350 of isoform 1 have been described. Transcriptional activity of GLI-2 is regulated by proteolytic processing and degradation. Within the region used as immunogen, mouse GLI-2 shares 91% and 97% amino acid sequence homology with that of human and rat GLI-2, respectively.

Long Name

GLI-Kruppel family member GLI2

Alternate Names

GLI2, THP2

Entrez Gene IDs

2736 (Human); 14633 (Mouse); 304729 (Rat)

Gene Symbol

GLI2

UniProt

NP_001074594

Additional GLI-2 Products

Product Documents for Mouse GLI‑2 Antibody

Certificate of Analysis

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Product Specific Notices for Mouse GLI‑2 Antibody

For research use only

Citations for Mouse GLI‑2 Antibody

Customer Reviews for Mouse GLI‑2 Antibody (5)

3.4 out of 5
5 Customer Ratings
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Showing  1 - 5 of 5 reviews Showing All
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  • Mouse GLI-2 Antibody
    Name: Oliver Tkaczyk
    Application: Western Blot
    Sample Tested: NIH-3T3 mouse embryonic fibroblast cell line
    Species: Mouse
    Verified Customer | Posted 08/02/2020
    Mouse GLI‑2 Antibody AF3635
  • Mouse GLI-2 Antibody
    Name: Brygida Baran
    Application: Immunoprecipitation
    Sample Tested: NIH-3T3 mouse embryonic fibroblast cell line
    Species: Mouse
    Verified Customer | Posted 07/22/2020
    Mouse GLI‑2 Antibody AF3635
  • Mouse GLI-2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MEF (fibroblast)
    Species: Mouse
    Verified Customer | Posted 07/08/2019
    This antibody recognizes many bands, and the major expected bands are not stronger (or maybe much weaker) than those non-specific bands.
    Mouse GLI‑2 Antibody AF3635
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to hear that this antibody did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: NIH-3T3 cells
    Species: Mouse
    Verified Customer | Posted 11/13/2015
    Gli2 Western blot
    Mouse GLI‑2 Antibody AF3635
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: NIH-3T3 cells
    Species: Mouse
    Verified Customer | Posted 11/13/2015
    Immunofluorescence staining of Gli2
    Mouse GLI‑2 Antibody AF3635

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