Shh, Recombinant, Mouse (Sonic HedgeHog), Highly Purified

Members of the Hedgehog (Hh) family are highly conserved proteins which are widely represented throughout the animal kingdom. The three known mammalian Hh proteins, Sonic (Shh), Desert (Dhh) and Indian (Ihh) are structurally related and share a high degree of amino-acid sequence identity (e.g., Shh and Ihh are 93% identical). The biologically active form of Hh molecules is obtained by autocatalytic cleavage of their precursor proteins and corresponds to approximately the N-terminal one half of the precursor molecule. Although Hh proteins have unique expression patterns and distinct biological roles within their respective regions of secretion, they use the same signaling pathway and can substitute for each other in experimental systems. Sonic Hedgehog, which is expressed only during embryogenesis, is instrumental in patterning the early embryo. It has been implicated as the key inductive signal in patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. Of three human proteins showing sequence and functional similarity to the Sonic Hedgehog protein of Drosophila, this protein is the most similar. The protein is made as a precursor that is autocatalytically cleaved, the N-terminal portion is soluble and contains the signalling activity while the C-terminal portion is involved in precursor processing. More importantly, the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product, restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. Defects in this protein or in its signalling pathway are a cause of holoprosencephaly (HPE), a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres. HPE is manifested by facial deformities. In addition, it is thought that mutations in this gene or in its signalling pathway may be responsible for VACTERL syndrome, which is characterized by vertebral defects, anal atresia, tracheoesophageal fistula with esophageal atresia, radial and renal dysplasia, cardiac anomalies, and limb abnormalities. , Source: DNA sequence encoding amino acid residues Cys 25-Gly 198 of mouse Shh (Echelard, Y. et al., 1993, Cell 75:1417-1430) was fused to a 6X histidine tag at the carboxy-terminus. The fusion protein was expressed in E. coli. Activity: Measured by its ability to induce alkaline phosphatase production by C3H10T1/2 fibroblasts (Nakamura, T. et al., 1997, Biochem. Biophys. Res. Commun. 237:465). The ED50 for this effect is typically 0.6-3ug/ml. , Endotoxin: < 1EU/ug, Molecular Mass: The 180 amino acid residue recombinant mouse Shh-N has a predicted molecular mass of approximately 20kD. , Reconstitution: We recommend a quick spin followed by reconstitution in sterile PBS, >0.1% BSA or HSA to a concentration of 0.05-1mg/ml. This solution can be diluted into other aqueous buffers and stored at 4°C for 1 week or -20°C for future use. Storage and Stability: Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS, >0.1% BSA or HSA, aliquot and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.