Recombinant Human Dkk-1 GMP Protein, CF

  
  • Purity
    >95%, by SDS-PAGE with silver staining.
  • Endotoxin Level
    <1.0 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to inhibit Wnt induced TCF reporter activity in HEK293 human embryonic kidney cells. Recombinant Human Dkk-1 (Catalog # 5439-GMP) inhibits a constant dose of 500 ng/mL of Recombinant Human Wnt-3a (Catalog # 5036‑WN). The ED50 for this effect is 10-60 ng/mL.
  • Source
    Spodoptera frugiperda, Sf 21 (baculovirus)-derived Thr32-His266 Manufactured and tested under cGMP guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis

    Thr-Leu-Asn-Ser-Val-Leu-Asn-Ser-Asn-Ala

  • Predicted Molecular Mass
    25.8 kDa
  • SDS-PAGE
    33-38 kDa, reducing conditions
5439-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. 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 6 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
Recombinant Human Wnt-3a (Catalog # 5036-WN) induces a dose responsive increase in Wnt reporter activity in HEK293 cells (green circles). Recombinant Human Dkk-1 GMP inhibits a constant dose of 500 ng/mL of Recombinant Human Wnt-3a. The ED50 for this effect is 10-60 ng/mL (orange circles).

1 μg/lane of Recombinant Human GMP-grade Dkk-1 was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a band at 34 kDa.

Background: Dkk-1

Dickkopf related protein 1 (Dkk-1) is the founding member of the Dickkopf family of proteins that includes Dkk-1, -2, -3, -4, and a related protein, Soggy (1, 2). Dkk proteins are secreted proteins that contain two conserved cysteine-rich domains separated by a linker region. Each domain contains ten cysteine residues (1‑3). Mature human Dkk-1 is a 40 kDa glycosylated protein that shares 86%, 87%, 90% and 91% aa sequence identity with mouse, rat, rabbit and bovine Dkk-1, respectively. It also shares 42% and 36% aa identity with human Dkk-2 and Dkk-4, respectively. Dkk-1 and Dkk-4 are well documented antagonists of the canonical Wnt signaling pathway (1, 2). This pathway is activated by Wnt engagement of a receptor complex composed of the Frizzled proteins and one of two low-density lipoprotein receptor-related proteins, LRP5 or LRP6 (4). Dkk-1 antagonizes Wnt by forming ternary complexes of LRP5/6 with Kremen1 or Kremen2 (4, 5). Dkk‑1/LRP6/Krm2 complex internalization has been shown to downregulate Wnt signaling (4, 5). Dkk-1 is expressed throughout development and antagonizes Wnt-7a during limb development (6, 7). Other sites of expression include developing neurons, hair follicles and the retina of the eye (8, 9). The balance between Wnt signaling and Dkk-1 inhibition is critical for bone formation and homeostasis (10). Insufficient or excess Dkk-1 activity in bone results in increased or decreased bone density, respectively (8, 11). In adults, Dkk-1 is expressed in osteoblasts and osteocytes, and neurons. Cerebral ischemia induces Dkk-1 expression, which contributes to neuronal cell death (12).

  • References:
    1. Krupnik, V.E. et al. (1999) Gene 238:301.  
    2. Niehrs, C. (2006) Oncogene 25:7469. 
    3. Bullock, C.M. et al. (2004) Mol. Pharmacol. 65:582. 
    4. Mao, B. et al. (2001) Nature 411:321. 
    5. Mao, B. et al. (2002) Nature 417:664.  
    6. Kemp, C. et al. (2005) Dev. Dyn. 233:1064.
    7. Adamska, M. et al. (2004) Dev. Biol. 272:134.
    8. Li, J. et al. (2006) Bone 36:754.
    9. Verani, R. et al. (2006) J. Neurochem. 101:242.
    10. Pinzone, J.J. et al. (2009) Blood 113:517.
    11. Morvan, F. et al. (2006) J. Bone Miner. Res. 21:934.
    12. Cappuccio, I. et al. (2005) J. Neurosci. 25:2647.
  • Long Name:
    Dickkopf-1
  • Entrez Gene IDs:
    22943 (Human); 13380 (Mouse)
  • Alternate Names:
    dickkopf (Xenopus laevis) homolog 1; dickkopf homolog 1 (Xenopus laevis); dickkopf related protein-1; Dickkopf-1; dickkopf-related protein 1; Dkk1; Dkk-1; hDkk-1; SKdickkopf-1 like

GMP Proteins
R&D Systems' 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 9001:2008 and ISO 13485:2003 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)

Additional testing and documentation requested by the customer can be arranged at an additional cost. Testing may include, but is not limited to, USP sterility 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.

 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.

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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.

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TERMS AND CONDITIONS
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