Recombinant Mouse Dkk-4 Protein Summary
Product Specifications
Optimal concentrations should be determined by each laboratory for each application. Measured by its ability to inhibit Wnt induced TCF reporter activity in HEK293 human embryonic kidney cells. Recombinant Mouse Dkk-4 (Catalog # 3105-DK) inhibits a constant dose of 100 ng/mL of Recombinant Mouse Wnt‑3a (Catalog # 1324-WN). The ED50 for this effect is 0.2-1.2 µg/mL.
Leu19-Ile221 (Arg67Ser and Arg70Ser), with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
3105-DK
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
| 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.
|
3105-DK/CF
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 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.
|
Scientific Data
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Recombinant Mouse Wnt-3a (Catalog # 1324-WN) induces a dose responsive increase in Wnt reporter activity in HEK293 cells (green circles). Recombinant Mouse Dkk-4 (Catalog # 3105-DK) inhibits a constant dose of 100 ng/mL of Recombinant Mouse Wnt-3a. The ED50 for this effect is 0.2-1.2 µg/mL (orange circles).
Reconstitution Calculator
Background: Dkk-4
Dickkopf related protein 4 (Dkk-4) is a member of the Dickkopf protein family that includes Dkk-1, -2, -3, and -4 and a related protein, Soggy (1). Expression of Dkk-4 has only been shown with sensitive PCR techniques during early embryonic development in mice (2), in differentiated human ES cells (3), or in mice that express dominant-active beta -catenin elevating Wnt signaling in the forebrain (4). Dkk proteins are secreted proteins that are synthesized as precursors with an N-terminal signal peptide; all have two conserved cysteine-rich domains separated by a linker region which contains a potential furin type proteolytic cleavage site. The domains contain 10 cysteines each; prokineticin and colipase families show a configuration of cysteines similar to the second motif (5). Mouse Dkk-4 shows 91%, 72% and 71% amino acid (aa) identity with rat, human and canine Dkk-4, respectively, and 40-46% aa identity with other mouse Dkk proteins. Dkk-4 is predicted as a 25 kDa protein, but transfection of 293T cells produces a shorter (15-17 kDa) form containing only the second cysteine-rich domain, as well as longer (30-32 and 40 kDa) forms that do not appear to be glycosylated or form covalent multimers (1). Of the four Dkk proteins, Dkk-4 is the most like Dkk-1. Both are unequivocal antagonists of the canonical Wnt signaling pathway (1, 6), which is activated by Wnt protein engagement of a receptor complex composed of the Frizzled proteins and one of two low-density lipoprotein receptor-related proteins, LRP5 or LRP6 (7). Dkk-1 and Dkk-4 antagonize Wnt by direct high-affinity binding to LRP5/6, forming ternary complexes of LRP5/6 with the Kremens protein Krm2. Internalization of the complex is triggered, making LRP5/6 unavailable for interaction with Wnt ligands (6-9).
- Krupnik, V.E. et al. (1999) Gene 238:301.
- Kemp, C. et al. (2005) Dev. Dyn. 233:1064.
- Katoh, Y. and M. Katoh (2005) Int. J. Mol. Med. 16:477.
- Diep, D.B. et al. (2004) Dev. Brain Res. 153:261.
- Bullock, C.M. et al. (2004) Mol. Pharmacol. 65:582.
- Mao, B. and C. Niehrs (2003) Gene 302:179.
- Logan, C.Y. and R. Nusse (2004) Ann. Rev. Cell Dev. Biol. 20:781.
- Mao, B. et al. (2002) Nature 417:664.
- Mao, B. et al. (2001) Nature 411:321.
Citation for Recombinant Mouse Dkk-4 Protein
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
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Identification of dkk4 as a target of Eda-A1/Edar pathway reveals an unexpected role of ectodysplasin as inhibitor of Wnt signalling in ectodermal placodes.
Authors: Fliniaux I, Mikkola ML, Lefebvre S, Thesleff I
Dev. Biol., 2008-04-26;320(1):60-71.
Species: Mouse
Sample Types: Whole Tissue
Applications: Bioassay
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