Overview

  • Product nameAnti-Gli3 antibodySee all Gli3 primary antibodies ...
  • Description
    Rabbit polyclonal to Gli3
  • Tested applicationsWB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Chimpanzee, Rhesus monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Gli3.

    (Peptide available as ab69917.)

  • Positive control
    • This antibody gave a positive signal in the following Lysates: Mouse Pancreas Tissue, Mouse Liver Tissue, MEF1 Whole Cell, TE 671 Whole Cell, RAW 264.7 Whole Cell

Properties

Applications

Our Abpromise guarantee covers the use of ab69838 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 145 kDa (predicted molecular weight: 160 kDa).Can be blocked with Human Gli3 peptide (ab69917).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionHas a dual function as a transcriptional activator and a repressor of the sonic hedgehog (Shh) pathway, and plays a role in limb development. The full-length GLI3 form (GLI3FL) after phosphorylation and nuclear translocation, acts as an activator (GLI3A) while GLI3R, its C-terminally truncated form, acts as a repressor. A proper balance between the GLI3 activator and the repressor GLI3R, rather than the repressor gradient itself or the activator/repressor ratio gradient, specifies limb digit number and identity. In concert with TRPS1, plays a role in regulating the size of the zone of distal chondrocytes, in restricting the zone of PTHLH expression in distal cells and in activating chondrocyte proliferation. Binds to the minimal GLI-consensus sequence 5'-GGGTGGTC-3'.
  • Tissue specificityIs expressed in a wide variety of normal adult tissues, including lung, colon, spleen, placenta, testis, and myometrium.
  • Involvement in diseaseDefects in GLI3 are the cause of Greig cephalo-poly-syndactyly syndrome (GCPS) [MIM:175700]. GCPS is an autosomal dominant disorder affecting limb and craniofacial development. It is characterized by pre- and postaxial polydactyly, syndactyly of fingers and toes, macrocephaly and hypertelorism.
    Defects in GLI3 are a cause of Pallister-Hall syndrome (PHS) [MIM:146510]. PHS is characterized by a wide range of clinical manifestations. It mainly associates central or postaxial polydactyly, syndactyly, and hypothalamic hamartoma. Malformations are frequent in the viscera, e.g. anal atresia, bifid uvula, congenital heart malformations, pulmonary or renal dysplasia. It is an autosomal dominant disorder.
    Defects in GLI3 are a cause of type A1/B postaxial polydactyly (PAPA1/PAPB) [MIM:174200, 603596]. PAPA in humans is an autosomal dominant trait characterized by an extra digit in the ulnar and/or fibular side of the upper and/or lower extremities. The extra digit is well formed and articulates with the fifth, or extra, metacarpal/metatarsal, and thus it is usually functional.
    Defects in GLI3 are a cause of polydactyly preaxial type 4 (POP4) [MIM:174700]. Polydactyly preaxial type 4 (i.e., polydactyly on the radial/tibial side of the hand/foot) covers a heterogeneous group of entities. In preaxial polydactyly type IV, the thumb shows only the mildest degree of duplication, and syndactyly of various degrees affects fingers 3 and 4.
    Defects in GLI3 are the cause of acrocallosal syndrome (ACS) [MIM:200990]; also abbreviated ACLS. ACS is characterized by postaxial polydactyly, hallux duplication, macrocephaly, and absence of the corpus callosum, usually with severe developmental delay.
  • Sequence similaritiesBelongs to the GLI C2H2-type zinc-finger protein family.
    Contains 5 C2H2-type zinc fingers.
  • Post-translational
    modifications
    Phosphorylated on multiple sites by protein kinase A (PKA) and phosphorylation by PKA primes further phosphorylation by CK1 and GSK3. Phosphorylation is essential for its proteolytic processing.
    Transcriptional repressor GLI3R, a C-terminally truncated form, is generated from the full-length GLI3 protein (GLI3FL/GLI3-190) through proteolytic processing. This process requires PKA-primed phosphorylation of GLI3, ubiquitination of GLI3 and the presence of BTRC. GLI3FL is complexed with SUFU in the cytoplasm and is maintained in a neutral state. Without the Hh signal, the SUFU-GLI3 complex is recruited to cilia, leading to the efficient processing of GLI3FL into GLI3R. GLI3R formation leads to its dissociation from SUFU, allowing it to translocate into the nucleus, and repress Hh target genes. When Hh signaling is initiated, SUFU dissociates from GLI3FL and this has two consequences. First, GLI3R production is halted. Second, free GLI3FL translocates to the nucleus, where it is phosphorylated, destabilized, and converted to a transcriptional activator (GLI3A). Phosphorylated in vitro by ULK3.
  • Cellular localizationNucleus. Cytoplasm. Cell projection > cilium. GLI3FL is localized predominantly in the cytoplasm while GLI3R resides mainly in the nucleus. Ciliary accumulation requires the presence of KIF7 and SMO. Translocation to the nucleus is promoted by interaction with ZIC1.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACLS antibody
    • DNA binding protein antibody
    • GCPS antibody
    • Gli 3 antibody
    • GLI family zinc finger 3 antibody
    • GLI Kruppel family member GLI 3 antibody
    • GLI Kruppel family member GLI3 (Greig cephalopolysyndactyly syndrome) antibody
    • GLI Kruppel family member GLI3 antibody
    • GLI3 antibody
    • GLI3 C-terminally truncated form antibody
    • GLI3 form of 190 kDa antibody
    • GLI3 form of 83 kDa antibody
    • GLI3 full length protein antibody
    • GLI3-190 antibody
    • GLI3-83 antibody
    • GLI3_HUMAN antibody
    • GLI3FL antibody
    • Glioma associated oncogene family zinc finger 3 antibody
    • Oncogene GLI3 antibody
    • PAP A antibody
    • PAPA 1 antibody
    • PAPA antibody
    • PAPA1 antibody
    • PAPB antibody
    • PHS antibody
    • PPD IV antibody
    • PPDIV antibody
    • Transcriptional activator GLI3 antibody
    • Transcriptional repressor GLI3R antibody
    • Zinc finger protein GLI 3 antibody
    • Zinc finger protein GLI3 antibody
    see all

Anti-Gli3 antibody images

  • All lanes : Anti-Gli3 antibody (ab69838) at 1 µg/ml

    Lane 1 : Pancreas (Mouse) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 4 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
    Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 160 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 110 kDa,55 kDa,75 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab69838 stained RAW 246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab69838, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) RAW 246.7 cells at 5µg/ml.

References for Anti-Gli3 antibody (ab69838)

This product has been referenced in:
  • Dormoy V  et al. Vitamin D3 triggers antitumor activity through targeting hedgehog signaling in human renal cell carcinoma. Carcinogenesis : (2012). Read more (PubMed: 22843547) »

See 1 Publication for this product

Product Wall

Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (8% acrylamide gel)
Sample Rat Tissue lysate - whole (Liver)
Specification Liver
Treatment CCl4
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Jan 09 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (AGS)
Specification AGS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Aug 15 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Placenta)
Specification Placenta
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
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Submitted Nov 17 2010

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