Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF GMP
R&D Systems, part of Bio-Techne | Catalog # 1314-GMP
Animal-Free
Key Product Details
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.
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 20425 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.
The ED50 for this effect is <5 μg/mL.
Host Cell Protein
<0.5 ng per µg of protein when tested by ELISA.
Mycoplasma
Negative for Mycoplasma.
Host Cell DNA
<0.0015 ng per µg of protein when tested by PCR.
Scientific Data Images for Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF
Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP Bioactivity
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 ED50 for this effect is <5 ug/mL.
Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP SDS-PAGE
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
1314-GMP
| 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.
|
Background: Sonic Hedgehog/Shh
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.
Alternate Names
HHG1, HLP3, HPE3, MCOPCB5, Shh, ShhNC, SMMCI, TPTPS
Gene Symbol
SHH
UniProt
Additional Sonic Hedgehog/Shh Products
Product Documents for Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF
Manufacturing Specifications
GMP ProteinsR&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.
R&D Systems' quality focus includes:
- Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
- Documented processes and QA control of documentation and process changes
- Personnel training programs
- Raw material testing and vendor qualification/monitoring
- Fully validated equipment, processes and test methods
- Equipment calibration schedules using a computerized calibration program
- Facility maintenance, safety programs and pest control
- Material review process for variances
- Monitoring of stability over product shelf-life
- N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
- Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
- Host Cell Protein testing performed by ELISA
- Mycoplasma testing by ribosomal RNA hybridization assay
Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.
R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.
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.
- 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.
- Low Endotoxin Level.
- No impairment of biological activity.
- High quality product obtained under stringent conditions.
Product Specific Notices for Recombinant Human Sonic Hedgehog/Shh, N-Terminus GMP, CF
END USER TERMS OF USE OF PRODUCT
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For preclinical, or clinical ex vivo use
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