Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

Name: Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free
Brand: R&D Systems
SKU: BT-SHH-AFL

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

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Product Datasheet / COA / SDS

Widely used for both human and mouse cell culture.

Key Product Details

Learn more about Animal-Free Recombinant Proteins

Source:	E. coli
Accession #:	Q62226.2
Applications:	Bioactivity

View Full Technical Details for Recombinant Mouse Sonic Hedgehog/Shh Protein, Animal-Free

Technical Data
Citations / Reviews

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

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.
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	19.8 kDa
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 Examples 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

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.

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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

Item	Value
References	Briscoe, J. and P.P. Therond (2013) Mol. Cell. Biol. 14:416. Aviles, E.C. et al. (2013) Front. Cell. Neurosci. 7:86. Xie, J. et al. (2013) OncoTargets Ther. 6:1425. Echelard, Y. et al. (1993) Cell 75:1417. Zeng, X. et al. (2001) Nature 411:716. Feng, J. et al. (2004) Development 131:4357. Goetz, J.A. et al. (2006) J. Biol. Chem. 281:4087. Pepinsky, R.B. et al. (1998) J. Biol. Chem. 273:14037. Chen, M.-H. et al. (2004) Genes Dev. 18:641. Etheridge, L.A. et al. (2010) Development 137:133. Jakobs, P. et al. (2014) J. Cell Sci. 127:1726. Dierker, T. et al. (2009) J. Biol. Chem. 284:8013. Lewis, P.M. et al. (2001) Cell 105:599. Carpenter, D. et al. (1998) Proc. Natl. Acad. Sci. USA 95:13630. Filmus, J. and M. Capurro (2014) Matrix Biol. 35:248. Chuang, P.-T. and A.P. McMahon (1999) Nature 397:617.
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

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

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