Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab6050 in the following tested applications.

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

Application Notes
ICC/IF Use a concentration of 1 µg/ml. See Abreview.
WB Use at an assay dependent concentration. Predicted molecular weight: 170 kDa. PubMed: 23293081
IHC-Fr 1/100.

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.
  • Target information above from: UniProt accession P10071 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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



  • Predicted band size : 170 kDa


    Western blot for Gli3 antibody (ab6050) at 1/500 tested on the following tissue lysates:

    Lane 1 : Human Brain
    Lane 2:  Human Lung
    Lane 3:  Human Spleen
    Lane 4:  Mouse Brain
    Lane 5:  Mouse Lung

    Secondary ab: Goat anti-rabbit IgG ab6721 (1/5000)
    Exposure time: 3 minutes
    Expected molecular weight: 2 isoforms of Gli3 exist, one is the full length 170-190kDa and the other is a truncated isoform at ~80kDa. 
    Cell lysates were loaded at 20µg per lane.

    NB: This image shows the truncated isoform of Gli3 is detected by ab6050 in human lung (we do not know the identity of the 120-130kDa band).  It was not possible to detect Gli3 immunoreactivity by WB in the other lysates tested. This is likely due to low expression/abundance of Gli3 in these tissue lysates. Furthermore, Gli3 expression is likely to be developmentally regulated and induced, making it difficult to succesfully detect in whole tissue homogenates. The staining on the WB shows low background and minimal cross reactivity in whole homogenates. The 50kDa band is likely due to non-specific binding of secondary antibody.

  • Gil3 expression in the radial glia of the developing mouse neocortex. ab6050 Rabbit polyclonal to Gil3 on Frozen sections of mouse E12.5 whole brain, showing staing of the radial glia, of the developing neocortex. Sections were paraformaldehyde fixed, prior to heat mediated antigen retrieval in citric acid, and 14 hours incubation with ab6050 (1/100).

    See Abreview

  • ab6050 at 0.625 µg/ml staining human glioblastoma.

  • ICC/IF image of ab6050 stained HepG2 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 (ab6050, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

References for Anti-Gli3 antibody (ab6050)

This product has been referenced in:
  • McIntyre BA  et al. Gli3-mediated hedgehog inhibition in human pluripotent stem cells initiates and augments developmental programming of adult hematopoiesis. Blood 121:1543-52 (2013). WB, ICC/IF ; Human . Read more (PubMed: 23293081) »
  • Ma X  et al. Trophic and proliferative effects of Shh on motor neurons in embryonic spinal cord culture from wildtype and G93A SOD1 mice. BMC Neurosci 14:119 (2013). Read more (PubMed: 24119209) »

See all 4 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: .05% · Temperature: 21°C
Sample Chicken Cell (Retina)
Specification Retina
Permeabilization No
Fixative Paraformaldehyde
Username

Mr. Chris Zelinka

Verified customer

Submitted Sep 02 2013

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (E12.5 Whole brain section)
Specification E12.5 Whole brain section
Fixative Paraformaldehyde
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5%
Username

Dr. Dean Griffiths

Verified customer

Submitted Jan 05 2007

Thank you for your enquiry. ab6050 is a fast track antiserum which is an antibody that is yet to be fully characterized and sold at a reduced rate of £ 125.00 for 100 µg. I would be very willing to provide you with the antiserum at a reduced ra...

Read More
Abreviews
Application Western blot
Sample Mouse Tissue lysate - whole (brain)
Specification brain
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10%
Username

Miss. Tian Yu

Verified customer

Submitted Sep 15 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"