详细说明
Species Reactivity
Human, Mouse
Specificity
Detects human and mouse Sonic Hedgehog/Shh N‑Terminus in direct ELISAs and Western blots. In direct ELISAs, less than
15% cross‑reactivity with recombinant mouse (rm) Dhh N‑Terminus and rmIhh N-Terminus is observed.Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant mouse Sonic Hedgehog/Shh N‑Terminus
Cys25-Gly198 (Lys122Arg)
Accession # Q62226Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Western Blot | Detection of Human and Mouse Sonic Hedgehog/Shh by Western Blot. Western blot shows lysates of human stomach tissue and mouse stomach tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Sonic Hedgehog/Shh N‑Terminus Antigen Affinity-purified Polyclonal Antibody (Catalog # AF464) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # ). A specific band was detected for Sonic Hedgehog/Shh at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Immunohistochemistry | Sonic Hedgehog/Shh in Mouse Embryo. Sonic Hedgehog/Shh was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Goat Anti-Human/Mouse Sonic Hedgehog/Shh N-Terminus Antigen Affinity-purified Polyclonal Antibody (Catalog # AF464) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # ) and counterstained with hematoxylin (blue). Specific staining was localized to developing brain. View our protocol for . |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Sonic Hedgehog/Shh
The hedgehog (hh) gene encoding a secreted protein was originally identified in Drosophila as a segment polarity gene. The vertebrate homologues of Hh comprise several proteins including sonic hedgehog (Shh), Indian hedgehog (Ihh), and Desert hedgehog (Dhh). Hedgehog proteins are important signaling molecules during embryonic development. Shh genes are highly conserved and have been identified in a variety of species including human, mouse, frog, fish, and chicken. Mouse and human Shh are 92% identical at the amino acid sequence level. Shh is expressed in key embryonic tissues such as the Hensen’s node, the zone of polarizing activity in the posterior limb bud, the notochord, and the floor plate of the neural tube. Shh is involved in regulating the patterning of the developing central nervous system, somite, and limb. Shh plays an important role in the development of particular tissues such as whisker, hair, foregut, tooth and bone. Evidence also suggests that Shh is involved in regulating stem cell fates of neural and hematopoeitic lineages, and that aberrant Shh signaling is implicated in basal cell carcinomas and other diseases.
Mouse Shh cDNA encodes a 437 amino acid residue with a predicted 24 aa residue signal peptide that is cleaved to generate a 413 aa residue precursor protein. An autocatalytic reaction yields a 19 kDa amino-terminal domain Shh-N protein containing cholesterol and palmitate, and a 27 kDa carboxy-terminal domain Shh-C protein. The N-terminal domain retains all known signaling capabilities, while the C-terminal domain is responsible for the intramolecular processing, acting as a cholesterol transferase. Shh can act as both a short-range contact dependent factor and as a long-range, diffusible morphogen. At the cell surface, Shh activity is mediated by a multicomponent receptor complex involving the 12-pass transmembrane protein Patched (Ptc) which binds Shh with high affinity and Smoothened (Smo), a signaling seven transmembrane G-protein coupled receptor. In the absence of Shh, Ptc represses Smo activity. The binding of Shh to Ptc, releases the basal repression of Smo by Ptc (1‑5).
References:
Carpenter, D. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13630.
Perrimon, N. (1995) Cell 80:517.
Weed, M. et al. (1997) Matrix Biol. 16:53.
Mullor, J. et al. (2002) Trends Cell Biol. 12:562.
Ingham, P. and A. McMahon (2001) Genes & Dev. 15:3059.
Entrez Gene IDs:
6469 (Human); 20423 (Mouse)
Alternate Names:
HHG1; HHG-1; HLP3; HPE3; MCOPCB5sonic hedgehog (Drosophila) homolog; Shh; SMMCIsonic hedgehog homolog (Drosophila); sonic hedgehog homolog; sonic hedgehog protein; Sonic Hedgehog; TPT; TPTPS