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AF3690

GLI-3 Antibody

(3 Reviews)

(100 Publications)

100 μg

AF3690

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.

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 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.

Conditional deletion of GATA6 does not significantly alter Gli3 processing in the hindlimb bud.(A) Western blot analysis of Gli3FL and Gli3R steady state levels in anterior (lanes 1 and 3) or posterior (lanes 2 and 4) halves of E11.5 hindlimb buds isolated from either control (lanes 1 and 2) or Prx1-Cre; GATA6fl/fl (lanes 3 and 4) mouse embryos. alpha –Tubulin is used as a loading control. (B) Quantification of Gli3FL/Gli3R ratios (by densitometry of Western Blots) in anterior or posterior halves of E11.5 hindlimb buds isolated from either littermate controls (lanes 1 and 2) or Prx1-Cre; GATA6fl/fl (lanes 3 and 4) animals, 12 embryos of each genotype were analyzed. Error bar indicates standard deviation, n = 12. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24415953), licensed under a CC-BY license. Not internally tested by R&D Systems.

SufuT396I does not stabilize Gli3FL protein and reduces the processing of Gli3FL.(A) Western blotting of lysates prepared from SufuR146X/R146X at E9.5 and SufuT396I/T396I, SufuT396I/+, and wild-type embryos at E10.5 with anti-Gli3, anti-Sufu, and anti-actin antibodies. Each image presented in the Fig. is a representative of independent triplicated experiments. The full gel images are shown in S6A Fig. (B, C) Relative expression of Gli3FL (B) and Gli3REP (B and C). Western blotting was performed two times using lysates prepared from five wild-type and five SufuT396I/T396I embryos at E10.5 (S2A Fig.). Expression levels were quantified from the band intensity shown in S2A Fig. as relative values of the Gli3FL/actin and Gli3REP/actin expression ratios (B) and the direct ratio of Gli3REP/Gli3FL (C). (**) p < 0.01, two-tailed Student’s t-test. Error bars indicate the standard deviations. (D) Western blotting of cell lysates from wild-type and SufuT396I/T396I MEFs with indicated antibodies. The SufuT396I/T396I MEFs were electroporated with 10.0, 1.0, or 0.1 μg of the HA–Sufu construct (lane 3, 4, and 5, respectively), or 10 μg of the HA–SufuT396I construct (lane 6). The mobilities on SDS-PAGE of the wild-type Sufu (lane 3–5) and SufuT396I (lane 6) are identical. The complete gel images are shown in S6B Fig. This image is representative of two independent experiments. (E) Western blotting of cell lysates from Sufu−/− cells with indicated antibodies. The Sufu−/− cells were electroporated with a mixture of the Flag–Gli3 construct (6 μg) and the HA–Sufu construct (4.00, 1.33, 0.44, or 0.15 μg) for the wild-type Sufu cotransfection (lane 2 to 5, respectively), or a mixture of the Flag–Gli3 construct (6 μg) and the HA–SufuT396I construct (4 μg) for mutant Sufu cotransfection (lane 6). The complete gel images are shown in S6C Fig. This image is representative of two independent experiments. (F) Western blotting of immunoprecipitates or lysates from 293T cells transfected with expression constructs as indicated at the top. The complete gel images are shown in S6D Fig. This image is representative of two independent experiments. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25760946), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, ,

Applications: , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF3690

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF3690-SP

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MAB3690

GLI-3 Antibody (2352B) [Unconjugated]

100 μg

MAB3690

GLI‑3 was detected in immersion fixed SW480 fibroblast carcinoma cell line (positive staining) and A549 human lung carcinoma cell line (negative staining) using Rabbit Anti-Human GLI‑3 Monoclonal Antibody (Catalog # MAB3690) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB3690-SP

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BAF3690

GLI-3 Antibody [Biotin]

50 μg

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

Applications:

Host:

Goat Polyclonal

Host: Goat Polyclonal

Conjugate:

Biotin

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF3690
Carrier Free	AF3690-SP

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