Recombinant Human Sonic Hedgehog/Shh (C24II) Animal-Free, CF

Name: Recombinant Human Sonic Hedgehog/Shh (C24II) Animal-Free, CF
Brand: R&D Systems
SKU: AFL1845
Price: 393 USD

Catalog #: AFL1845 Datasheet / COA / SDS

GMP Version Available: 1845-GMP

GMP

Intended for preclinical researchers who may transition to GMP Sonic Hedgehog/Shh (C24II) for their clinical work

Catalog #	Availability	Size / Price	Qty
AFL1845-025
AFL1845-100
AFL1845-500

Recombinant Human Sonic Hedgehog/SHH (C23IL) Animal-Free Bioactivty.

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All Sonic Hedgehog/Shh Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Sonic Hedgehog/Shh (C24II) Animal-Free, CF Summary

Learn more about Animal-Free Recombinant Proteins

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

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 human Sonic Hedgehog/Shh protein
Cys24-Gly197 (Cys24Ile-Ile), with an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.

Accession #

NP_000184.1

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

20 kDa

SDS-PAGE

22 kDa, reducing conditions

Product Datasheets

AFL1845

Product Datasheet

COA

SDS

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.

AFL1845

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS and NaCl.
Reconstitution	Reconstitute at 100-200 μg/mL in PBS, and allow up to 24 hours at 2 to 8 °C for complete reconstitution.
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.

Scientific Data

Bioactivity

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Animal-Free™ Recombinant Human Sonic Hedgehog/Shh (Catalog # AFL1845) 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.

SDS-PAGE

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1 μg/lane of Animal-Free™ Recombinant Human Sonic Hedgehog/Shh (C24II) Protein (Catalog # AFL1845) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 22 kDa.

Reconstitution Calculator

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

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

HHG1; HHG-1; HLP3; HPE3; MCOPCB5; MCOPCB5sonic hedgehog (Drosophila) homolog; Shh; ShhNC; SMMCI; SMMCIsonic hedgehog homolog (Drosophila); sonic hedgehog homolog; sonic hedgehog protein; Sonic Hedgehog; TPT; TPTPS

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