Recombinant Human Sonic Hedgehog/Shh (C24II) N-Term GMP, CF

Animal-Free.
  
  • Purity
    >95%, by SDS-PAGE with silver staining, under reducing conditions.
  • Endotoxin Level
    <0.01 EU per 1 μg of the protein by the LAL method.
  • 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 0.1-0.4 µg/mL.
  • Source
    E. coli-derived Cys24-Gly197 (Cys24Ile-Ile), with an N-terminal Met Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under current Good Manufacturing Practice (GMP) guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis
    Met-Ile-Ile-Gly25-Pro-Gly-Arg-Gly-Phe-Gly
  • Predicted Molecular Mass
    20 kDa
  • SDS-PAGE
    22 kDa, reducing conditions
1845-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in PBS and NaCl.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images

GMP-grade Recombinant Human Sonic Hedgehog/Shh (C24II), N-Terminus (Catalog # 1845-GMP) induces alkaline phosphatase production by the C3H10T1/2 mouse embryonic fibroblast cell line. The ED50 for this effect is 0.1-0.4 μg/mL.

1 μg/lane of GMP-grade Recombinant Human Sonic Hedgehog/Shh (C24II)
N-Term (Catalog # 1845-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing R band at 21.5 kDa and NR band at 22.5 kDa, respectively.
Mass Spectrometry
LC/ESI-MS analysis of GMP-grade Recombinant Human Sonic Hedgehog/Shh (C24II) N-Terminus (Catalog # 1845-GMP). The major peak at 19809 corresponds to the calculated molecular mass, 19814 Da.
Background: Sonic Hedgehog/Shh
Sonic Hedgehog (Shh) is expressed in embryonic tissues that are critical for the patterning of the developing central nervous system, somite, and limb. It is also involved in whisker, hair, foregut, tooth, and bone development. Shh regulates neural and hematopoietic stem cell fate and is important for thymocyte differentiation and proliferation as well as T cell determination. In adult tissue Shh is associated with cancer development and tissue remodeling following injury (1-3). Human Shh encodes a 462 amino acid (aa) precursor protein that is autocatalytically processed to yield a non-glycosylated 19 kDa N-terminal fragment (Shh-N) and a glycosylated 25 kDa C-terminal protein (Shh-C) (4). Shh-C, which is responsible for the intramolecular processing of Shh, is rapidly degraded following Shh proteolysis (5). Shh-N is highly conserved, sharing >98% aa identity between mouse, human, rat, canine, porcine, and chicken Shh-N. Shh-N can be palmitoylated at its
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).
  • References:
    1. Briscoe, J. and P.P. Therond (2013) Mol. Cell. Biol. 14:416.
    2. Aviles, E.C. et al. (2013) Front. Cell. Neurosci. 7:86.
    3. Xie, J. et al. (2013) OncoTargets Ther. 6:1425.
    4. Marigo, V. et al. (1995) Genomics 28:44.
    5. Zeng, X. et al. (2001) Nature 411:716.
    6. Feng, J. et al. (2004) Development 131:4357.
    7. Goetz, J.A. et al. (2006) J. Biol. Chem. 281:4087.
    8. Pepinsky, R.B. et al. (1998) J. Biol. Chem. 273:14037.
    9. Chen, M.-H. et al. (2004) Genes Dev. 18:641.
    10. Etheridge, L.A. et al. (2010) Development 137:133.
    11. Jakobs, P. et al. (2014) J. Cell Sci. 127:1726.
    12. Dierker, T. et al. (2009) J. Biol. Chem. 284:8013.
    13. Lewis, P.M. et al. (2001) Cell 105:599.
    14. Carpenter, D. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13630.
    15. Filmus, J. and M. Capurro (2014) Matrix Biol. 35:248.
    16. Chuang, P.-T. and A.P. McMahon (1999) Nature 397:617.
  • 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

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