WB, IP
H
Transfected Only
85
Rabbit IgG
#Q99835
6608
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:200 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human Smoothened protein.
Background
Smoothened is a 7-pass transmembrane G protein-coupled receptor that functions as a key transmembrane transducer of Hedgehog signaling (1). The primary function of Smoothened is to regulate the activity of GLI-family transcription factors (GLI1-3). In the canonical (mammalian) Hedgehog signaling pathway, Smoothened activity is constitutively repressed by the 12-pass transmembrane receptor Patched (PTCH1). PTCH1 represses Smoothened activity by controlling the availability of cholesterol, which serves as a second messenger agonist for Smoothened (2). This repression results in the association of GLI proteins with the STK36/SUFU protein complex, promoting GLI protein cleavage and nuclear translocation of “repressor” GLI protein fragments that suppress target gene transcription (1). Binding of Hedgehog ligands inactivates the PTCH1 receptor, resulting in the derepression of Smoothened. Active Smoothened promotes the release of GLI proteins from the STK36/SUFU protein complex, enabling cytoplasmic-to-nuclear translocation of full-length GLI proteins that activate target gene expression. The Hedgehog signaling pathway regulates cell fate, embryonic patterning, and postnatal tissue repair, but is generally quiescent in healthy postnatal tissues. However, research studies have shown that ectopic activation of the pathway (e.g., through aberrant Smoothened activity) can be oncogenic, making Smoothened and other components of the Hedgehog signaling pathway attractive targets for therapeutic intervention in some cancer types (3-5).
- Ingham, P.W. et al. (2011) Nat Rev Genet 12, 393-406.
- Radhakrishnan, A. et al. (2020) Nat Chem Biol 16, 1303-1313.
- Barakat, M.T. et al. (2010) Trends Mol Med 16, 337-48.
- Briscoe, J. and Thérond, P.P. (2013) Nat Rev Mol Cell Biol 14, 416-29.
- Jeng, K.S. et al. (2020) Int J Mol Sci 21, 6863. doi: 10.3390/ijms21186863.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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