Recombinant Human Sonic Hedgehog/Shh Protein, High Activity

R&D Systems, part of Bio-Techne | Catalog # 8908-SH

C-terminal cholesterol, N-terminal fatty acid-modified
R&D Systems, part of Bio-Techne
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Carrier Free
8908-SH-005/CF
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With Carrier
8908-SH-005
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Key Product Details

Source

HEK293

Accession #

Q15465

Structure / Form

Cholesterol-modified at the C-terminal and fatty acid-modified at the N-terminal

Conjugate

Unconjugated

Applications

Bioactivity

View all Sonic Hedgehog/Shh Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human Sonic Hedgehog/Shh protein
Cys24-Gly197

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Cys24

Predicted Molecular Mass

20 kDa

SDS-PAGE

18-24 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 typically 6-36 ng/mL.

Reviewed Applications

Read 1 review rated 4 using 8908-SH in the following applications:

Scientific Data Images for Recombinant Human Sonic Hedgehog/Shh Protein, High Activity

Enhanced Activity of Human Cell-expressed Sonic Hedgehog (Shh) Protein.

Enhanced Activity of Human Cell-expressed Sonic Hedgehog (Shh) Protein.

Recombinant Human Shh proteins induce alkaline phosphatase production by mesenchymal stem cells. High Activity Shh (green), purified from HEK293 cells and containing the correct post-translational modifications (cholesterol and fatty acids), is over 14-fold more active than E. coli-purified Recombinant Human Shh-N (C24II) N-Terminus (Catalog # 1845-SH; red line), and over 250-fold more active than E. coli-purified Recombinant Human Shh-N (Catalog # 1314-SH; blue line).
Enhanced Activity of Human Cell-expressed Sonic Hedgehog (Shh) Protein.

Enhanced Activity of Human Cell-expressed Sonic Hedgehog (Shh) Protein.

1 μg/lane of Recombinant Human Sonic Hedgehog/Shh Protein was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing 18-24 kDa bands.
Post-translational Modification Analysis of Naturally-modified Recombinant Human Sonic Hedgehog (Shh) Protein.

Post-translational Modification Analysis of Naturally-modified Recombinant Human Sonic Hedgehog (Shh) Protein.

LC/ESI-MS analysis of Recombinant Human (rh)SHH Protein, High Activity shows major peaks at 20119.3, 20145.2, and 20171.6 Da, suggesting that recombinant human SHH molecules are dual-modified with cholesterol at C-terminus, and fatty acids (lauric acid, myristic acid, and palmitic acid) at the N-terminus. The minor peaks at 19776 Da corresponds to rhSHH with only fatty acid modification.

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: 8908-SH
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and CHAPS with BSA as a carrier protein.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Carrier Free: 8908-SH/CF
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and CHAPS.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Sonic Hedgehog/Shh

The Sonic Hedgehog (Shh) protein 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. The Shh protein 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 Shh 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). The Shh-C protein, which is responsible for the intramolecular processing of Shh, is rapidly degraded following Shh proteolysis (5). The Shh-N protein 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 the Shh protein as well as its assembly into various sized multimers (6-9). Lipid modification and multimerization greatly increase the receptor binding affinity and signaling potency of the Shh-N protein (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 functionality of the Shh protein and are required for the development of Shh protein gradients important in tissue morphogenesis (9, 13). Canonical signaling by the Shh protein is mediated by a multicomponent receptor complex that includes Patched (PTCH1, PTCH2) and Smoothened (SMO) (14). Binding of the Shh protein 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.

Alternate Names

HHG1, HLP3, HPE3, MCOPCB5, Shh, ShhNC, SMMCI, TPTPS

Entrez Gene IDs

6469 (Human); 20423 (Mouse)

Gene Symbol

SHH

UniProt

Q15465

Additional Sonic Hedgehog/Shh Products

Product Documents for Recombinant Human Sonic Hedgehog/Shh Protein, High Activity

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Recombinant Human Sonic Hedgehog/Shh Protein, High Activity

For research use only

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