Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
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.
|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-P||Use a concentration of 0.5 - 5 µg/ml.|
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.
modificationsPhosphorylated 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
- ACLS antibody
- DNA binding protein antibody
- GCPS antibody
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 20
µg 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, makin
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).
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.
This product has been referenced in:
- Chen Y et al. Generation of iPSC-derived limb progenitor-like cells for stimulating phalange regeneration in the adult mouse. Cell Discov 3:17046 (2017). Read more (PubMed: 29263795) »
- Ma X et al. Crosstalk between Notch and Sonic hedgehog signaling in a mouse model of amyotrophic lateral sclerosis. Neuroreport 28:141-148 (2017). Read more (PubMed: 27984541) »