Recombinant Human Dkk-1 GMP Protein, CF

Name: Recombinant Human Dkk-1 GMP Protein, CF
Brand: R&D Systems
SKU: 5439-GMP

Catalog #: 5439-GMP Datasheet / COA / SDS

Discontinued Product

5439-GMP has been discontinued.

View all Dkk-1 products.

Recombinant Human Dkk-1 GMP Protein Bioactivity

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All Dkk-1 Proteins and Enzymes

Product Details

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

Recombinant Human Dkk-1 GMP Protein, CF Summary

Product Specifications

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 human Dkk-1 protein
Thr32-His266
Manufactured and tested under cGMP guidelines.

Accession #

O94907

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

Product Datasheets

5439-GMP

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.

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 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.

Scientific Data

Bioactivity

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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).

SDS-PAGE

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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.

Reconstitution Calculator

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

Krupnik, V.E. et al. (1999) Gene 238:301.
Niehrs, C. (2006) Oncogene 25:7469.
Bullock, C.M. et al. (2004) Mol. Pharmacol. 65:582.
Mao, B. et al. (2001) Nature 411:321.
Mao, B. et al. (2002) Nature 417:664.
Kemp, C. et al. (2005) Dev. Dyn. 233:1064.
Adamska, M. et al. (2004) Dev. Biol. 272:134.
Li, J. et al. (2006) Bone 36:754.
Verani, R. et al. (2006) J. Neurochem. 101:242.
Pinzone, J.J. et al. (2009) Blood 113:517.
Morvan, F. et al. (2006) J. Bone Miner. Res. 21:934.
Cappuccio, I. et al. (2005) J. Neurosci. 25:2647.

Long Name

Dickkopf-1

Entrez Gene IDs

22943 (Human); 13380 (Mouse); 102123735 (Cynomolgus Monkey)

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

Manufacturing Specifications

GMP Proteins
R&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:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

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 application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, 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.