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Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CFGMP


Order Information
Key Product Details
Source: | E. coli |
Accession #: | Q15465 |
Applications: | Bioactivity |
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Product Specifications
Source |
E. coli-derived human Sonic Hedgehog/Shh protein Cys24-Gly197, with a C-terminal 6-His tag Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines. |
Purity | >97%, by SDS-PAGE with silver staining, under reducing conditions. Intact mass analysis of recombinant human sonic hedgehog confirms the predicted molecular mass of 20383 Da |
Endotoxin level | <0.10 EU per 1 μg of the protein by the LAL method. |
N-terminal sequence Analysis | (Cys24)-Gly-Pro-Gly-Arg-Gly-Phe-Gly-Lys-Arg |
Predicted Molecular Mass | 20 kDa |
SDS-PAGE | 22 kDa, reducing conditions |
Activity | Measured by its ability to induce alkaline phosphatase production by C3H10T1/2 mouse embryonic fibroblast cells. Nakamura, T. et al. (1997) Biochem. Biophys. Res. Commun. 237:465. The ED50 for this effect is <5 μg/mL. |
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GMP-grade Recombinant Human Sonic Hedgehog/Shh, N-Terminus (Catalog # 1314-GMP) induces alkaline phosphatase production by the C3H10T1/2 mouse embryonic fibroblast cell line. The ED50for this effect is <5 ug/mL. |
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1 μg/lane of GMP-grade Recombinant Human Sonic Hedgehog/Shh, N-Terminus (Catalog # 1314-GMP) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 22 kDa. |
Formulation, Preparation and Storage
Carrier Free
What does CF mean?CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
What formulation is right for me?In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
Carrier Free
Formulation | Lyophilized from a 0.2 μm filtered solution in NaH2PO4, NaCl and DTT. |
Reconstitution | Reconstitute at 250 μg/mL in sterile deionized water. |
Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Background: Sonic Hedgehog/Shh
N-terminal cysteine and modified by cholesterol addition at its C-terminus (6). These modifications contribute to the membrane tethering of Shh as well as its assembly into various sized multimers (6-9). Lipid modification and multimerization greatly increase Shh-N receptor binding affinity and signaling potency (5, 6, 8, 9). Monomeric and multimeric Shh can be released from the plasma membrane by the cooperative action of DISP1, SCUBE2, and TACE/ADAM17 (10-12). Modifications also extend the effective range of Shh functionality and are required for the development of protein gradients important in tissue morphogenesis (9, 13). Canonical signaling of Shh is mediated by a multicomponent receptor complex that includes Patched (PTCH1, PTCH2) and Smoothened (SMO) (14). The binding of Shh to PTCH releases the basal repression of SMO by PTCH. Shh activity can also be regulated through interactions with heparin, glypicans, and membrane-associated Hip (hedgehog interacting protein) (13, 15, 16).
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References |
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Entrez Gene IDs | 6469 (Human); 20423 (Mouse) |
Alternate Names | HHG-1, HHG1, HLP3, HPE3, MCOPCB5, MCOPCB5sonic hedgehog (Drosophila) homolog, SMMCI, SMMCIsonic hedgehog homolog (Drosophila), Shh, ShhNC, Sonic Hedgehog, TPT, TPTPS, sonic hedgehog homolog, sonic hedgehog protein |
Citations
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