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

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  • 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.
  • 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.
  • 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.
  • Accession #
  • 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
Product Datasheets

Certificate of Analysis Lookup

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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.
  • 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.
 
Data Images
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.
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.
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:
    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. Marigo, V. et al. (1995) Genomics 28:44.
    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.
  • Entrez Gene IDs:
    6469 (Human); 20423 (Mouse)
  • Alternate Names:
    HHG1; HHG-1; HLP3; HPE3; MCOPCB5sonic hedgehog (Drosophila) homolog; Shh; SMMCIsonic hedgehog homolog (Drosophila); sonic hedgehog homolog; sonic hedgehog protein; Sonic Hedgehog; TPT; TPTPS
Manufacturing Specifications
GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.

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

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

  • 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 bioburden testing (using broth culture, Sabourand's dextrose and blood agar plates with results reported at 3 days and at 7 days)
  • Host Cell Protein testing performed by ELISA
  • Mycoplasma testing by ribosomal RNA hybridization assay

Additional testing and documentation requested by the customer can be arranged at an additional cost. Testing may include, but is not limited to, USP <61> bioburden testing, positive identity testing, testing for adventitious agents and testing for residual host cell content.

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 its GMP grade recombinant protein products for research use or further manufacturing use in ex vivo cell therapy applications. They are not for in vivo use or for use as therapeutic or other drugs, biologic products or devices. 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.

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

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells its GMP products for research use only or further manufacturing and not for in vivo use, the production of therapeutics or other drugs or for biologic products or devices. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

 

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The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

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Primary Antibodies
Description Application Cat# Citations Images  

His Tag Antibody

WB , Simple Western , Flow , AP , CyTOF-ready MAB050 28  
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His Tag HRP-conjugated Antibody

WB , Simple Western MAB050H 3  
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His Tag APC-conjugated Antibody

ICFlow IC050A  
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His Tag PE-conjugated Antibody

ICFlow IC050P 3  
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His Tag Biotinylated Antibody

WB BAM050 4  
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His Tag Antibody

WB , Flow , AP MAB050R 2  
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
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His Tag PerCP-conjugated Antibody

ICFlow IC050C  
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Protein Purification Kits
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Histidine-tagged Protein Purification Resin

IP999 1
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