Description
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 and are highly conserved within and across species. Mouse and human Ihh share 100% amino acid identity in the signaling domain, while mouse Ihh and Shh share 90% amino acid identity in the N-terminal signaling domain. Ihh mRNA expression is detected in fetal lung, gut, stomach, liver, kidney, pancreas and strongly in cartilage - in growth regions of the developing bone. Ihh, along with parathyroid hormone related protein, regulate the rate of chondrocyte proliferation and differentiation. Ihh is also involved in yolk sac vasculogenesis, playing an important role in differentiation of epiblast cells into endothelial and red blood cells.Mouse Ihh cDNA encodes a 411 amino acid (aa) polypeptide with a predicted 27 aa signal peptide. This polypeptide is cleaved to generate a 45 kDa precursor protein that undergoes the same post‑translation processing as Shh. An autocatalytic reaction yields a 19 kDa amino‑terminal domain Ihh‑N protein that retains all known signaling capabilities, and a 23 kDa carboxy‑terminal domain Ihh-C protein. Since hydrophobic modifications to Shh, including the substitution of the N-terminal cysteine residue with two hydrophobic isoleucine residues, can also increase its potency, a similar modification was made for Ihh. This modified form also shows increased potency in a bioassay measuring induction of alkaline phosphatase. At the cell surface, Hedgehog activity is mediated by a multicomponent receptor complex involving the 12‑pass transmembrane protein Patched (Ptc) which binds Hedgehogs with high affinity and Smoothened (Smo), a signaling seven transmembrane G-protein coupled receptor.
Molecular Weight
Approximately 19.8 kDa.
AA sequence
IIGPGRVVGS RRRPPRKLVP LAYKQFSPNV PEKTLGASGR YEGKIARSSE RFKELTPNYN PDIIFKDEEN TGADRLMTQR CKDRLNSLAI SVMNQWPGVK LRVTEGWDED GHHSEESLHY EGRAVDITTS DRDRNKYGLL ARLAVEAGFD WVYYESKAHV HCSVKSEHSA AAKTGG
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
> 96 % by SDS-PAGE and HPLC analyses.
Biological Activity
The ED50 as determined by its ability to induce alkaline phosphatase production by C3H10T1/2(CCL-226) cells is 3.0-10 μg/mL. Fully biologically active when compared to standard.
Endotoxin
< 1.0 EU per 1μg of the protein by the LAL method.
Formulation
Lyophilized from a 0.2 µm filtered concentrated solution in 1 × PBS, pH 7.4.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20℃. Further dilutions should be made in appropriate buffered solutions.
