Human Dkk-2 Antibody

Catalog # Availability Size / Price Qty
MAB6628-100
MAB6628-SP
Dkk‑2  in SH-SY5Y Human Cell Line.
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Human Dkk-2 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Dkk-2 in direct ELISAs.
Source
Monoclonal Mouse IgG2A Clone # 994930
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Chinese Hamster Ovary cell line, CHO-derived human Dkk-2
Met1-Ile259
Accession # NP_055236
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Immunohistochemistry
5-25 µg/mL
See below
Immunocytochemistry
5-25 µg/mL
See below
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Immunocytochemistry Dkk‑2  in SH-SY5Y Human Cell Line. View Larger

Dkk‑2 in SH-SY5Y Human Cell Line. Dkk‑2 was detected in immersion fixed SH‑SY5Y human neuroblastoma cell line using Mouse Anti-Human Dkk‑2 Monoclonal Antibody (Catalog # MAB6628) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Dkk‑2  in Human Kidney. View Larger

Dkk‑2 in Human Kidney. Dkk‑2 was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human Dkk‑2 Monoclonal Antibody (Catalog # MAB6628) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Intracellular Staining by Flow Cytometry Detection of DKK-2 in Human SHSY-5Y Cells by Flow Cytometry View Larger

Detection of DKK-2 in Human SHSY-5Y Cells by Flow Cytometry Human SHSY-5Y neuroblastoma cell line was stained with Mouse Anti-Human DKK-2 Monoclonal Antibody (Catalog # MAB6628, filled histogram) or Mouse IgG2A Isotype Control Antibody (Catalog # MAB003, open histogram) followed by Goat anti-Mouse IgG PE-conjugated Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Dkk-2

Dickkopf related protein 2 (Dkk-2) is a member of the Dickkopf family of secreted Wnt modulators (1-3). Dkk proteins contain a signal peptide and two conserved cysteine-rich domains that are separated by a linker region. The second cysteine-rich domain mediates Dkk-2 binding activities, and its interaction with  beta -propeller domains of LRP‑5/6 has been mapped (2-4, 7). The 226 amino acid (aa), ~35 kDa mature human Dkk-2 shares 96%, 97%, 97%, 97%, 97% and 98% aa identity with mouse, rat, canine, equine, bovine and porcine Dkk-2, respectively. Mouse Dkk-2 can activate the canonical Wnt signaling pathway in Xenopus embryos, showing evolutionary conservation of function (5). Dkk proteins modify Wnt engagement of a receptor complex composed of a Frizzled protein and a low-density lipoprotein receptor-related protein, either LRP‑5 or LRP‑6 (3). Also, Kremen-1 and Kremen-2 are high affinity receptors for Dkk-1 and Dkk-2 (9). When LRP‑6 is over-expressed, direct high‑affinity binding of Dkk-2 to LRP can enhance canonical Wnt signaling (6-8). However, when Dkk‑2 and LRP‑6 form a ternary complex with Kremen‑2, Wnt signaling is inhibited due to internalization of Dkk‑2/LRP6/Krm2 complexes (9, 10). Thus, depending on the cellular context, Dkk‑2 can either activate or inhibit canonical Wnt signaling (3). In contrast, binding of Dkk-1 or Dkk-4 to LRP is consistently antagonistic (3). Dkk proteins are expressed in mesenchymal tissues and control epithelial transformations. Dkk-2 expression has been studied most in bone and eye, although it is expressed as early as periimplantation in mice (11). Mouse Dkk-1 or Dkk-2 deficiencies have opposite effects on bone homeostasis, despite down‑regulating Wnt antagonism in both cases (12, 13). 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 (12). In contrast, Dkk-1 negatively regulates late osteoblast proliferation, which limits bone density (13). Dkk-2-deficient mice are blind, exhibiting faulty differentiation of corneal epithelium and ectopic blood vessels in the periocular mesenchyme (14, 15).

References
  1. Monaghan, A.P. et al. (1999) Mech. Dev. 87:45.
  2. Krupnik, V.E. et al. (1999) Gene 238:301.
  3. Niehrs, C. (2006) Oncogene 25:7469.
  4. Chen, L. et al. (2008) J. Biol. Chem. 283:23364.
  5. Wu, W. et al. (2000) Current Biol. 10:1611.
  6. Mao, B. et al. (2001) Nature 411:321.
  7. Li, L. et al. (2002) J. Biol. Chem. 277:5977.
  8. Brott, B. and S.Y. Sokol (2002) Mol. Cell. Biol. 22:6100.
  9. Mao, B. et al. (2002) Nature 417:664.
  10. Mao, B. and C. Niehrs (2003) Gene 302:179.
  11. Zhang, Y. et al. (2009) J. Reprod. Dev. 55:17.
  12. Li, X. et al. (2005) Nat. Genet. 37:945.
  13. van der Horst, G. et al. (2005) J. Bone Miner. Res. 20:1867.
  14. Mukhopadhyay, M. et al. (2006) Development 133:2149.
  15. Gage, P.J. et al. (2008) Dev. Biol. 317:310.
Long Name
Dickkopf-2
Entrez Gene IDs
27123 (Human); 56811 (Mouse)
Alternate Names
dickkopf (Xenopus laevis) homolog 2; dickkopf 2; dickkopf homolog 2 (Xenopus laevis); dickkopf related protein-2; dickkopf-2; dickkopf-related protein 2; Dkk2; Dkk-2; hDkk-2

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FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Mouse IgG2A Isotype Control

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Reconstitution Buffers

Secondary Antibodies

Mouse IgG2A Antibody

Mouse IgG2A Antibody

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