Human Dkk-1 Quantikine ELISA Kit

Product is in the process of discontinuation and replaced by catalog # DKK100B.
(10 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 (10 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
  • Sensitivity
    15.6 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    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
QC241, Quantikine Immunoassay Control Set 921 for Human Dkk-1 - Please Inquire
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-free cell culture supernates, serum, and platelet-poor 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 natural human Dkk-1.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean22360013532215771287
Standard Deviation9.415.336.516.93569.6
CV%4.22.62.77.76.15.4

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean21259913582436201371
Standard Deviation8.924.544.418.437.463.1
CV%4.24.13.37.364.6

Cell Culture Supernates, Serum-free Cell Culture Supern, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean75.618736075.6192372
Standard Deviation2.213.289.485.8116.730.1
CV%2.91.82.67.78.78.1

Recovery

The recovery of human 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
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Dkk-1 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Human 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
      Dickkopf-1
    • 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.
    Citations:

    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.

    10 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Correlation of blood bone turnover biomarkers and Wnt signaling antagonists with AS, DISH, OPLL, and OYL
      Authors: CC Niu, SS Lin, LJ Yuan, LH Chen, CY Yang, AN Chung, ML Lu, TT Tsai, PL Lai, WJ Chen
      BMC Musculoskelet Disord, 2017;18(1):61.
      Species: Human
      Sample Type: Serum
    2. Interleukin-32 Gamma Stimulates Bone Formation by Increasing miR-29a in Osteoblastic Cells and Prevents the Development of Osteoporosis
      Authors: EJ Lee, SM Kim, B Choi, EY Kim, YH Chung, EJ Lee, B Yoo, CK Lee, S Hong, BJ Kim, JM Koh, SH Kim, YG Kim, EJ Chang
      Sci Rep, 2017;7(0):40240.
      Species: Human
      Sample Type: Plasma
    3. Effects of exercise training on circulating levels of Dickkpof-1 and secreted frizzled-related protein-1 in breast cancer survivors: A pilot single-blind randomized controlled trial
      Authors: TH Kim, JS Chang, KS Park, J Park, N Kim, JI Lee, ID Kong
      PLoS ONE, 2017;12(2):e0171771.
      Species: Human
      Sample Type: Serum
    4. MicroRNA-433-3p promotes osteoblast differentiation through targeting DKK1 expression
      Authors: X Tang, J Lin, G Wang, J Lu
      PLoS ONE, 2017;12(6):e0179860.
      Species: Rat
      Sample Type: Serum
    5. Gene expression profiling identifies microphthalmia-associated transcription factor (MITF) and Dickkopf-1 (DKK1) as regulators of microenvironment-driven alterations in melanoma phenotype.
      Authors: Hartman M, Talar B, Noman M, Gajos-Michniewicz A, Chouaib S, Czyz M
      PLoS ONE, 2014;9(4):e95157.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Evidences of early senescence in multiple myeloma bone marrow mesenchymal stromal cells.
      Authors: Andre T, Meuleman N, Stamatopoulos B, De Bruyn C, Pieters K, Bron D, Lagneaux L
      PLoS ONE, 2013;8(3):e59756.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. 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
    8. 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.
    9. 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
    10. 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
    ELISA Controls
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    Quantikine Immunoassay Control Set 921 for Human Dkk-1

    QC241
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