Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

R&D Systems, part of Bio-Techne | Catalog # BT-SHH-AFL

Widely used for both human and mouse cell culture.
R&D Systems, part of Bio-Techne
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Key Product Details

Source

E. coli

Accession #

Q62226.2

Conjugate

Unconjugated

Applications

Bioactivity

View all Sonic Hedgehog/Shh Proteins and Enzymes »

Product Specifications

Source

E. coli-derived mouse Sonic Hedgehog/Shh protein
Cys25-Gly198 (Cys25Ile-Ile) with an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.

Purity

>97%, by SDS-PAGE with 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

Met

Predicted Molecular Mass

19.8 kDa

SDS-PAGE

21 kDa, under 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 0.0500-0.600 μg/mL.

Scientific Data Images for Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

Equivalent Bioactivity of GMP, Animal-Free, and RUO grades of Recombinant Mouse Sonic Hedgehog/Shh.

Equivalent bioactivity of GMP (BT-SHH-GMP), Animal-Free (Catalog # BT-SHH-AFL) and RUO grades of Recombinant Mouse Sonic Hedgehog/Shh as measured in the alkaline phosphatase production using C3H10T1/2 mouse embryonic fibroblast cells (orange, green, red, respectively).

Animal-Free™ Recombinant Mouse Sonic Hedgehog/Shh Protein Bioactivity.

Animal-Free™ Recombinant Mouse Sonic Hedgehog/Shh Protein (Catalog # BT-SHH-AFL) induces alkaline phosphatase production by the C3H10T1/2 mouse embryonic fibroblast cell line. The ED50 for this effect is 0.0500-0.600 μg/mL.

Animal-Free™ Recombinant Mouse Sonic Hedgehog/Shh Protein SDS-PAGE.

2 μg/lane of Animal-Free™ Recombinant Mouse Sonic Hedgehog/Shh Protein (Catalog # BT-SHH-AFL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at 21 kDa under reducing conditions.

Formulation, Preparation and Storage

BT-SHH-AFL
Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Phosphate, NaCl and DTT with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in 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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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). Mouse Shh encodes a 437 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. Echelard, Y. et al. (1993) Cell 75:1417.
  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

Q62226.2

Additional Sonic Hedgehog/Shh Products

Product Documents for Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.  

 Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

    Quality Assurance

    • Low Endotoxin Level.
    • No impairment of biological activity.
    • High quality product obtained under stringent conditions.
    • For ex vivo research or bioproduction, additional documentation can be provided.

    Please read our complete Animal-Free Statement

    Product Specific Notices for Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

    For research use or further manufacturing only

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