Human/Mouse GLI-3 Antibody

Catalog # Availability Size / Price Qty
AF3690
AF3690-SP
Detection of Mouse GLI‑3 by Western Blot.
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Product Details
Citations (28)
FAQs
Supplemental Products
Reviews (1)

Human/Mouse GLI-3 Antibody Summary

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

Western Blot Detection of Mouse GLI‑3 by Western Blot. View Larger

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. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human/Mouse GLI-3 Antibody

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.

28 Citations: Showing 1 - 10
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  1. Axoneme polyglutamylation regulated by Joubert syndrome protein ARL13B controls ciliary targeting of signaling molecules
    Authors: K He, X Ma, T Xu, Y Li, A Hodge, Q Zhang, J Torline, Y Huang, J Zhao, K Ling, J Hu
    Nat Commun, 2018;9(1):3310.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  2. The ciliary protein RPGRIP1L governs autophagy independently of its proteasome-regulating function at the ciliary base in mouse embryonic fibroblasts
    Authors: A Struchtrup, A Wiegering, B Stork, U Rüther, C Gerhardt
    Autophagy, 2018;0(0):1-47.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: WB
  3. Spatiotemporal manipulation of ciliary glutamylation reveals its roles in intraciliary trafficking and Hedgehog signaling
    Authors: SR Hong, CL Wang, YS Huang, YC Chang, YC Chang, GV Pusapati, CY Lin, N Hsu, HC Cheng, YC Chiang, WE Huang, NC Shaner, R Rohatgi, T Inoue, YC Lin
    Nat Commun, 2018;9(1):1732.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  4. Spop regulates Gli3 activity and Shh signaling in dorsoventral patterning of the mouse spinal cord
    Authors: H Cai, A Liu
    Dev. Biol., 2017;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: WB
  5. 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 Types: Cell Lysates
    Applications: WB
  6. 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 Types: Cell Lysates
    Applications: WB
  7. Regulation of Calvarial Osteogenesis by Concomitant De-repression of GLI3 and Activation of IHH Targets
    Authors: LK Veistinen, T Mustonen, MR Hasan, M Takatalo, Y Kobayashi, DA Kesper, A Vortkamp, DP Rice
    Front Physiol, 2017;8(0):1036.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: WB
  8. 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 Types: Cell Lysates
    Applications: WB
  9. EMT programs promote basal mammary stem cell and tumor-initiating cell stemness by inducing primary ciliogenesis and Hedgehog signaling
    Authors: VJ Guen, TE Chavarria, C Kröger, X Ye, RA Weinberg, JA Lees
    Proc. Natl. Acad. Sci. U.S.A., 2017;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  10. Gorlin syndrome-derived induced pluripotent stem cells are hypersensitive to hedgehog-mediated osteogenic induction
    Authors: D Hasegawa, H Ochiai-Shi, S Onodera, T Nakamura, A Saito, T Onda, K Watanabe, K Nishimura, M Ohtaka, M Nakanishi, K Kosaki, A Yamaguchi, T Shibahara, T Azuma
    PLoS ONE, 2017;12(10):e0186879.
  11. Mechanism of inhibition of the tumor suppressor Patched by Sonic Hedgehog
    Proc Natl Acad Sci USA, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: WB
  12. Cholesterol activates the G-protein coupled receptor Smoothened to promote morphogenetic signaling
    Elife, 2016;5(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: WB
  13. Spop promotes skeletal development and homeostasis by positively regulating Ihh signaling
    Proc. Natl. Acad. Sci. U.S.A, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  14. Craniofacial Ciliopathies Reveal Specific Requirements for GLI Proteins during Development of the Facial Midline
    PLoS Genet., 2016;12(11):e1006351.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: WB
  15. 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 Types: Whole Tissue
    Applications: IHC - Paraffin embedded
  16. 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 Types: Cell Lysates
    Applications: WB
  17. Sufu and Kif7 in limb patterning and development.
    Authors: Zhulyn O, Hui C
    Dev Dyn, 2015;244(3):468-78.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: WB
  18. 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 Types: Cell Lysates
    Applications: WB
  19. 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 Types: Tissue Homogenates
    Applications: WB
  20. 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 Types: Cell Lysates
    Applications: WB
  21. 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 Types: Tissue Homogenates
    Applications: WB
  22. 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 Types: Cell Lysates
    Applications: WB
  23. 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 Types: Tissue Homogenates
    Applications: WB
  24. 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 Types: Cell Lysates
    Applications: WB
  25. 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.
  26. 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 Types: Whole Tissue
    Applications: IHC Paraffin-embedded
  27. 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 Types: Tissue Homogenates
    Applications: WB
  28. 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 Types: Cell Lysates
    Applications: IP

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Human/Mouse GLI‑3 Antibody
By Anonymous on 10/26/2015
Application: WB Sample Tested: Mouse embryo cell lysate Species: Mouse

Specificity: Specific
Sensitivity: Sensitive
Buffer: TBST
Dilution: 1/1000