Dkk-2, Recombinant, Mouse (Dickkopf-2, Dickkopf-related Protein 2 Precursor, Dkk-2, hDkk-2, UNQ682/PRO1316)

Dickkopf related protein 2 (Dkk-2) is a member of the Dickkopf family of secreted Wnt modulators. Dkk proteins contain a signal peptide and two conserved cysteine-rich domains that are separated by a linker region. The second cysteine-rich domain, which shows a configuration of cysteines conserved in prokineticin and colipase families, mediates Dkk-2 binding activities. \n The 226 amino acid (aa), ~35kD mature mouse Dkk-2 shares 41% and 34% aa identity with mouse Dkk-1 and Dkk-4, respectively. It also shares 99%, 96%, 96%, 96% and 94% aa \nidentity with rat, human, dog, horse and cow Dkk-2, respectively, and can activate the canonical Wnt signaling pathway in Xenopus embryos. Dkk proteins modify Wnt engagement of a receptor complex composed of a Frizzled protein and a low-density lipoprotein receptor-related \nprotein, either LRP5 or LRP6. When LRP6 is overexpressed, direct high-affinity binding of Dkk-2 to LRP can enhance canonical Wnt signaling. However, when Dkk-2 and LRP6 form a ternary complex with Kremen2, Wnt signaling is inhibited due to internalization of Dkk-2/LRP6/Krm2 complexes. Thus, depending on the cellular context, Dkk-2 can either activate or inhibit canonical Wnt signaling. Incontrast, binding of Dkk-1 or Dkk-4 to LRP is consistently antagonistic. \n\nDkk proteins are expressed in mesenchymal tissues and control epithelial transformations. Dkk-2 expression has been studied most in bone and eye. Mouse Dkk-1 or Dkk-2 deficiencies have opposite effects on bone homeostasis, despite downregulating Wnt antagonism in both cases. Dkk-2 expression is induced by Wnts in bone, and is thought to enhance bone density by promoting terminal differentiation of osteoblasts and mineral deposition. In contrast, Dkk-1 negatively regulates late osteoblast proliferation, which limits bone density. Dkk-2-deficient mice are blind due to faulty differentiation of corneal epithelium. \n\nMolecular Mass: \nBased on N-terminal amino acid sequencing, the recombinant mouse Dkk-2 starts at Ser 26 and has a calculated molecular mass of approximately 27.1kD. As a result of glycosylation, recombinant mouse Dkk-2 migrates as an approximately 30kD protein in SDS-PAGE under reducing conditions. \n\nWhen rmKremen-1 is coated at 4ug/ml (100ul/well), rmDkk-2 inhibits 50% binding of rhDkk-1 biotin (200 ng/mL) at the concentration range of 5-15ug/mL. Also measured by its ability to inhibit Wnt-3a induced alkaline phosphate synthesis in the presence of BMP-2 by MC3T3E1 (mouse pre-osteoblastic cells). \n\nEndotoxin: < 1.0 EU per 1ug of the receptor as determined by the LAL method. \n\nStorage and Stability:\n\nLyophilized powder may be stored at -20 degrees C. Reconstitute to nominal volume by adding sterile buffer. Aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

D3795-01

Size

10ug

Reactivities

Mouse

Form

Supplied as a lyophilized powder from PBS and BSA. Reconstitute with sterile PBS and 1% BSA or HSA.

Purity

Purity > 95%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin: < 1.0 EU per 1ug of the receptor as determined by the LAL method.

References

References: \n1. Monaghan, A.P. et al., 1999, Mech. Dev. 87:45. 8. Brott, B. & S.Y. Sokol, 2002, Mol. Cell. Biol. 22:6100. \n2. Krupnik, V.E. et al., 1999, Gene 238:301. 9. Mao, B. et al., 2002, Nature 417:664. \n3. Niehrs, C., 2006, Oncogene 25:7469. 10. Mao, B. & C. Niehrs, 2003, Gene 302:179. \n4. Bullock, C.M. et al., 2004, Mol. Pharmacol. 65:582. 11. Li, X. et al., 2005, Nat. Genet. 37:945. \n5. Wu, W. et al., 2000, Current Biol. 10:1611. 12. Morvan, F. et al., 2006, J. Bone Miner. Res. 21:934. \n6. Mao, B. et al., 2001, Nature 411:321. 13. Mukhopadhyay, M. et al., 2006, Development 133:2149. \n

Additional Info

Recognizes mouse Dkk-2. Species sequence homology: mouse Dkk-1: 41%, mouse Dkk-4: 34% ,rat: 99%, human Dkk-2: 96%, canine Dkk-2: 96%, horse Dkk-2: 96% and bovine Dkk-2: 94% It can also activate the canonical Wnt signaling pathway in Xenopus embryos.