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

Name: Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF
Brand: rnd
SKU: 1314-GMP

Catalog # 1314-GMP

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Animal-Free

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Key Product Details

Source:	E. coli
Accession #:	Q15465
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF

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.

Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF Scientific Data Examples

	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.
	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.

1314-GMP

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. 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.

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). 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).

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. Marigo, V. et al. (1995) Genomics 28:44. 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

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

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Product Specifications

Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF Scientific Data Examples

Formulation, Preparation and Storage

Carrier Free

Carrier Free

Reconstitution Calculator

Background: Sonic Hedgehog/Shh

Citations

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