Human/Mouse GLI-3 Antibody

(21 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse GLI-3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human (rh) GLI-1 and rhGLI-2 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human GLI-3
    Met1-Glu479
    Accession # P10071
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/106 cells
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse GLI‑3 by Western Blot. 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.
Chromatin Immunoprecipitation (ChIP)
Detection of GLI‑3-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 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.
Immunocytochemistry
GLI‑3 in HeLa Human Cell Line. 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: GLI-3
Human GLI-3 is a 190 kDa transcription factor of the C2H2-type zinc-finger protein family. GLI-3 appears to play a role in limb and brain development, probably through sonic hedgehog signaling. GLI-3 is regulated by PKA-mediated phosphorylation. Unphosphorylated full-length GLI-3 is a transcriptional activator, while phosphorylated GLI-3 is cleaved after amino acid 700 to generate an N-terminal 83 kDa transcriptional repressor. Over the region used as immunogen, human GLI-3 is 95% identical to the corresponding mouse and canine protein sequences.
  • Long Name:
    GLI-Kruppel family member GLI3
  • Entrez Gene IDs:
    2737 (Human); 14634 (Mouse); 140588 (Rat)
  • Alternate Names:
    ACLS; ACLSPAPAPHS; ADD; Bph; GCPS; GLI family zinc finger 3; GLI3 form of 190 kDa; GLI3 full length protein; GLI3; GLI-3; GLI3-190; GLI3FL; GLI-Kruppel family member GLI3; glioma-associated oncogene family zinc finger 3; Greig cephalopolysyndactyly syndrome; oncogene GLI3; PAP-A; PAPA1; PAPB; PAPBDNA-binding protein; Pdn; PHS; PPDIV; Xt; zinc finger protein GLI3
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

21 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Suppressor of Fused Restraint of Hedgehog Activity Level is Critical for Osteogenic Proliferation and Differentiation during Calvarial Bone Development
    Authors: J Li, Y Cui, J Xu, Q Wang, X Yang, Y Li, X Zhang, M Qiu, Z Zhang, Z Zhang
    J. Biol. Chem., 2017;0(0):.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  2. Regulation of Gli ciliary localization and Hedgehog signaling by the PY-NLS/karyopherin-?2 nuclear import system
    Authors: Y Han, Y Xiong, X Shi, J Wu, Y Zhao, J Jiang
    PLoS Biol., 2017;15(8):e2002063.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  3. Sonic Hedgehog Activates Phospholipase A2 to Enhance Smoothened Ciliary Translocation
    Authors: AM Arensdorf, ME Dillard, JM Menke, MW Frank, CO Rock, SK Ogden
    Cell Rep, 2017;19(10):2074-2087.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  4. Mechanism of inhibition of the tumor suppressor Patched by Sonic Hedgehog
    Proc Natl Acad Sci USA, 2016;0(0):.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  5. Spop promotes skeletal development and homeostasis by positively regulating Ihh signaling
    Proc. Natl. Acad. Sci. U.S.A, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Illegitimate translation causes unexpected gene expression from on-target out-of-frame alleles created by CRISPR-Cas9
    Sci Rep, 2016;6(0):39608.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  7. Cholesterol activates the G-protein coupled receptor Smoothened to promote morphogenetic signaling
    Elife, 2016;5(0):.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  8. Craniofacial Ciliopathies Reveal Specific Requirements for GLI Proteins during Development of the Facial Midline
    PLoS Genet., 2016;12(11):e1006351.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  9. T-box3 is a ciliary protein and regulates stability of the Gli3 transcription factor to control digit number
    Authors: U Emechebe, P Kumar P, JM Rozenberg, B Moore, A Firment, T Mirshahi, AM Moon
    Elife, 2016;5(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  10. Sufu and Kif7 in limb patterning and development.
    Authors: Zhulyn O, Hui C
    Dev Dyn, 2015;244(3):468-78.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  11. Bifurcating action of Smoothened in Hedgehog signaling is mediated by Dlg5.
    Authors: Chong Y, Mann R, Zhao C, Kato M, Beachy P
    Genes Dev, 2015;29(3):262-76.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  12. T396I mutation of mouse Sufu reduces the stability and activity of Gli3 repressor.
    Authors: Makino S, Zhulyn O, Mo R, Puviindran V, Zhang X, Murata T, Fukumura R, Ishitsuka Y, Kotaki H, Matsumaru D, Ishii S, Hui C, Gondo Y
    PLoS ONE, 2015;10(3):e0119455.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  13. Murine Joubert syndrome reveals Hedgehog signaling defects as a potential therapeutic target for nephronophthisis.
    Authors: Hynes, Ann Mari, Giles, Rachel H, Srivastava, Shalabh, Eley, Lorraine, Whitehead, Jennifer, Danilenko, Marina, Raman, Shreya, Slaats, Gisela G, Colville, John G, Ajzenberg, Henry, Kroes, Hester Y, Thelwall, Peter E, Simmons, Nicholas, Miles, Colin G, Sayer, John A
    Proc Natl Acad Sci U S A, 2014;111(27):9893-8.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  14. GATA6 is a crucial regulator of Shh in the limb bud.
    Authors: Kozhemyakina, Elena, Ionescu, Andreia, Lassar, Andrew B
    PLoS Genet, 2014;10(1):e1004072.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  15. A switch from low to high Shh activity regulates establishment of limb progenitors and signaling centers.
    Authors: Zhulyn O, Li D, Deimling S, Vakili N, Mo R, Puviindran V, Chen M, Chuang P, Hopyan S, Hui C
    Dev Cell, 2014;29(2):241-9.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  16. Ciliopathy proteins regulate paracrine signaling by modulating proteasomal degradation of mediators.
    Authors: Liu Y, Tsai I, Morleo M, Oh E, Leitch C, Massa F, Lee B, Parker D, Finley D, Zaghloul N, Franco B, Katsanis N
    J Clin Invest, 2014;124(5):2059-70.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  17. Regulation and function of immunosuppressive molecule human leukocyte antigen G5 in human bone tissue.
    Authors: Deschaseaux F, Gaillard J, Langonne A, Chauveau C, Naji A, Bouacida A, Rosset P, Heymann D, De Pinieux G, Rouas-Freiss N, Sensebe L
    FASEB J, 2013;27(8):2977-87.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  18. Gli3 regulation of myogenesis is necessary for ischemia-induced angiogenesis.
    Authors: Renault M, Vandierdonck S, Chapouly C, Yu Y, Qin G, Metras A, Couffinhal T, Losordo D, Yao Q, Reynaud A, Jaspard-Vinassa B, Belloc I, Desgranges C, Gadeau A
    Circ Res, 2013;113(10):1148-58.
  19. The hedgehog system machinery controls transforming growth factor-beta-dependent myofibroblastic differentiation in humans: involvement in idiopathic pulmonary fibrosis.
    Authors: Cigna N, Farrokhi Moshai E, Brayer S, Marchal-Somme J, Wemeau-Stervinou L, Fabre A, Mal H, Leseche G, Dehoux M, Soler P, Crestani B, Mailleux A
    Am J Pathol, 2012;181(6):2126-37.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  20. Prevention of premature fusion of calvarial suture in GLI-Kruppel family member 3 (Gli3)-deficient mice by removing one allele of Runt-related transcription factor 2 (Runx2).
    Authors: Tanimoto Y, Veistinen L, Alakurtti K
    J. Biol. Chem., 2012;287(25):21429-38.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  21. The output of Hedgehog signaling is controlled by the dynamic association between Suppressor of Fused and the Gli proteins.
    Authors: Humke EW, Dorn KV, Milenkovic L, Scott MP, Rohatgi R
    Genes Dev., 2010;24(7):670-82.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: IP
Expand to show all 21 Citations
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human/Mouse GLI‑3 Antibody AF3690
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 WB Mouse Anonymous 10/26/2015
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Western Blot Human/Mouse GLI‑3 Antibody AF3690
Western Blot: Human/Mouse GLI‑3 Antibody [AF3690]

Summary

ApplicationWestern Blot
Sample TestedMouse embryo cell lysate
SpeciesMouse

Other Experimental Details

Other Experimental DetailsSpecificity: Specific
Sensitivity: Sensitive
Buffer: TBST
Dilution: 1/1000

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