Human/Porcine Dkk-1 Quantikine ELISA Kit

(4 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (13 uL), EDTA Plasma (25 uL), Platelet-poor EDTA Plasma (100 uL), Heparin Plasma (25 uL), Platelet-poor Heparin Plasma (100 uL)
  • Sensitivity
    15.6 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor EDTA Plasma, Heparin Plasma, Platelet-poor Heparin Plasma)
  • Specificity
    Oligomeric forms of natural and recombinant human Dkk-1
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
Product Summary
The Quantikine Human Dkk-1 immunoassay is a 4.5 hour solid phase ELISA designed to measure human Dkk-1 in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human Dkk-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Dkk-1 showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of naturally occurring human Dkk-1.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Platelet-poor EDTA Plasma, Heparin Plasma, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation8.8524.544.418.437.463.1

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Standard Deviation9.3815.336.516.93569.6


The recovery of Dkk-1 spiked to various levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 97-109
EDTA Plasma (n=4) 94 86-102
Heparin Plasma (n=4) 98 92-106
Serum (n=4) 99 85-115
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of natural Dkk-1 were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human/Porcine Dkk-1 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Dkk-1
Members of the dickkopf-related protein family (Dkk-1, -2, -3, and -4) are secreted proteins with two cysteine-rich domains separated by a linker region. Dkk-3 and -4 also have one prokineticin domain. Dkk-1, -2, and -4 function as antagonists of canonical Wnt signaling by binding to LRP5/6, preventing interaction of LRP5/6 with Wnt-Frizzled complexes. Dkk-1, -2, and -4 also bind cell surface Kremen-1 or -2 and promote the internalization of LRP5/6. Antagonistic activity of Dkk-3 has not been demonstrated. Dkk proteins have distinct patterns of expression in adult and embryonic tissues and have a wide range of effects on tissue development and morphogenesis. The Dkk family also includes Soggy, which is homologous to Dkk-3 but not to the other family members. Soggy has not been shown to inhibit Wnt signaling, and its role in the pathway is unclear.
    • Long Name
    • 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;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    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.

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    Sample Type
    1. Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast.
      Authors: Scheel C, Eaton E, Li S, Chaffer C, Reinhardt F, Kah K, Bell G, Guo W, Rubin J, Richardson A, Weinberg R
      Cell, 2011;145(6):926-40.
    2. Pharmaceutical inhibition of glycogen synthetase kinase-3beta reduces multiple myeloma-induced bone disease in a novel murine plasmacytoma xenograft model.
      Authors: Gunn WG, Krause U, Lee N, Gregory CA
      Blood, 2011;117(5):1641-51.
      Species: Mouse
      Sample Type: Serum
    3. The roles of Wnt signaling modulators Dickkopf-1 (Dkk1) and Dickkopf-2 (Dkk2) and cell maturation state in osteogenesis on microstructured titanium surfaces.
      Authors: Olivares-Navarrete R, Hyzy S, Wieland M, Boyan BD, Schwartz Z
      Biomaterials, 2010;31(8):2015-24.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Dickkopf-1 (Dkk-1) in plasma and synovial fluid is inversely correlated with radiographic severity of knee osteoarthritis patients.
      Authors: Honsawek S, Tanavalee A, Yuktanandana P, Ngarmukos S, Saetan N, Tantavisut S
      BMC Musculoskelet Disord, 2010;11(0):257.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 736 for Human Dkk-1

    Ctrl QC76
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine Wash Buffer 1

    ELISA WA126 5
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