Hedgehog Signaling Antibody Sampler Kit #26118
Product Information
Kit Usage Information
Protocols
- 2207: Western Blotting
- 2468: Western Blotting, Immunoprecipitation (Agarose)
- 2470: Western Blotting, Immunoprecipitation (Magnetic)
- 2520: Western Blotting
- 3538: Western Blotting, Immunoprecipitation (Agarose)
- 7074: Western Blotting
- 18773: Western Blotting, Immunoprecipitation (Magnetic)
- 71107: Western Blotting, Immunoprecipitation (Magnetic)
- 92981: Western Blotting, Immunoprecipitation (Agarose)
Product Description
Specificity / Sensitivity
Shh (C9C5) Rabbit mAb detects endogenous levels of total Shh protein. This antibody does not cross-react with transfected IHH or DHH. PTCH1 (C53A3) Rabbit mAb detects transfected levels of PTCH1. This antibody can also detect endogenous levels of PTCH1 through immunoprecipitation followed by western blot analysis. PTCH2 (G1191) Antibody detects transfected levels of PTCH2 protein. It does not recognize transfected levels of human PTCH1 protein. Smoothened (E6Z5T) Rabbit mAb detects transfected levels of total smoothened (SMO) protein. SUFU (C54G2) Rabbit mAb detects endogenous levels of total SUFU protein. GLI1 (C68H3) Rabbit mAb detects endogenous levels of total GLI1 protein. GLI2 (E5V8N) Rabbit mAb detects endogenous levels of total GLI2 protein. GLI3 (E6E2K) Rabbit mAb detects endogenous levels of total GLI3 protein.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly1191 of human PTCH2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Glu53 of human Shh, Pro1307 of human PTCH1, Leu458 of human SUFU, Gly420 of human GLI1, and Pro1188 of human GLI2 protein, or recombinant proteins corresponding to human smoothened (SMO) protein, and the carboxy terminus of human GLI3.
Background
The Hedgehog (Hh) signaling pathway plays critical roles in the regulation of cell fate, tissue patterning, and growth during embryonic development. It is downregulated during postnatal development, but can be reactivated to promote tissue repair and regeneration. Aberrant Hh signaling activity during prenatal development is associated with numerous birth defects (e.g., holoprosencephaly), while uncontrolled Hh pathway activation postnatally is linked to the development of several cancer types (1,2). There are three canonical Hh ligands: Sonic hedgehog (SHH), Indian hedgehog (IHH), and Desert hedgehog (DHH), all of which have distinct as well as overlapping roles and expression patterns (3-5). Patched1 and 2 (PTCH1 and PTCH2) are partially redundant 12-pass transmembrane proteins that function as receptors for Hh ligands (6-8). Smoothened (SMO) is a 7-pass transmembrane G protein-coupled receptor (GPCR) that functions as the key transducer of Hh signaling. In the absence of Hh ligands (off-state), PTCH proteins are localized to cilia, and function to suppress SMO activity (1,2). Suppressor of Fused (SUFU) simultaneously contributes to suppression of the pathway by sequestering the glioma-associated oncogene (GLI) family of transcription factors (9,10). Binding of Hh ligands to PTCH receptors results in derepression of SMO, in part by promoting its translocation to cilia; this leads to downregulation of SUFU, resulting in the stabilization and nuclear translocation of GLI transcription factors that regulate the transcription of genes involved in cell proliferation, migration, and survival (1).
- Lee, R.T. et al. (2016) Development 143, 367-72.
- McMahon, A.P. et al. (2003) Curr Top Dev Biol 53, 1-114.
- Zhang, X.M. et al. (2001) Cell 106, 781-92.
- Adolphe, C. et al. (2004) Development 131, 5009-19.
- Pathi, S. et al. (2001) Mech Dev 106, 107-17.
- Stone, D.M. et al. (1996) Nature 384, 129-34.
- Motoyama, J. et al. (1998) Nat Genet 18, 104-6.
- Smyth, I. et al. (1999) Hum Mol Genet 8, 291-7.
- Barnfield, P.C. et al. (2005) Differentiation 73, 397-405.
- Méthot, N. and Basler, K. (2000) Development 127, 4001-10.
Limited Uses
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