Human Dkk-1 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1906
Ancillary Products Available

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Human Dkk-1 ELISA Standard Curve
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Citations (20)
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Human Dkk-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Dkk-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human Dkk-1 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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); 102123735 (Cynomolgus Monkey)
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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human Dkk-1 DuoSet ELISA

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.

20 Citations: Showing 1 - 10
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  1. Combination of Wnt/beta-Catenin Targets S100A4 and DKK1 Improves Prognosis of Human Colorectal Cancer
    Authors: M Dahlmann, A Monks, ED Harris, D Kobelt, M Osterland, F Khaireddin, P Herrmann, W Kemmner, S Burock, W Walther, RH Shoemaker, U Stein
    Cancers, 2021-12-22;14(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Effect of Dickkopf-1 (Dkk-1) and SP600125, a JNK Inhibitor, on Wnt Signaling in Canine Prostate Cancer Growth and Bone Metastases
    Authors: W Supsavhad, BB Hassan, JK Simmons, WP Dirksen, SM Elshafae, NA Kohart, AA Demirer, TJ Rosol
    Veterinary sciences, 2021-08-01;8(8):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Activity of fibroblast-like synoviocytes in rheumatoid arthritis was impaired by dickkopf-1 targeting siRNA
    Authors: YY Liu, SY Wang, YN Li, WJ Bian, LQ Zhang, YH Li, L Long, X Liu, XW Zhang, ZG Li
    Chin. Med. J., 2020-03-20;0(6):679-686.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Dickkopf-1 (DKK1) promotes tumor growth via Akt-phosphorylation and independently of Wnt-axis in Barrett's associated esophageal adenocarcinoma
    Authors: O Lyros, AK Lamprecht, L Nie, R Thieme, K Götzel, M Gasparri, G Haasler, P Rafiee, R Shaker, I Gockel
    Am J Cancer Res, 2019-02-01;9(2):330-346.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Three-dimensional in vitro modeling of malignant bone disease recapitulates experimentally accessible mechanisms of osteoinhibition
    Authors: EP McNeill, RW Reese, A Tondon, BH Clough, S Pan, J Froese, D Palmer, U Krause, DM Loeb, R Kaunas, CA Gregory
    Cell Death Dis, 2018-11-26;9(12):1161.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Dickkopf-related protein 1 and Gremlin 1 show different response than Frizzled-related protein in human synovial fluid following knee injury and in patients with osteoarthritis
    Authors: X Huang, JN Post, L Zhong, J Leijten, S Larsson, M Karperien, A Struglics
    Osteoarthr. Cartil., 2018-03-08;0(0):.
    Species: Human
    Sample Types: Synovial Fluid
  7. Exploring the Wnt signaling pathway in schizophrenia and bipolar disorder
    Authors: EZ Hoseth, F Krull, I Dieset, RH Mørch, S Hope, ES Gardsjord, NE Steen, I Melle, HR Brattbakk, VM Steen, P Aukrust, S Djurovic, OA Andreassen, T Ueland
    Transl Psychiatry, 2018-03-06;8(1):55.
    Species: Human
    Sample Types: Plasma
  8. Nitric Oxide Mediates Crosstalk between Interleukin 1? and WNT Signaling in Primary Human Chondrocytes by Reducing DKK1 and FRZB Expression
    Authors: L Zhong, S Schivo, X Huang, J Leijten, M Karperien, JN Post
    Int J Mol Sci, 2017-11-22;18(11):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Increase in Dickkopf-1 Serum Level in Recent Spondyloarthritis. Data from the DESIR Cohort.
    Authors: Nocturne G, Pavy S, Boudaoud S, Seror R, Goupille P, Chanson P, van der Heijde D, van Gaalen F, Berenbaum F, Mariette X, Briot K, Feydy A, Claudepierre P, Dieude P, Nithitham J, Taylor K, Criswell L, Dougados M, Roux C, Miceli-Richard C
    PLoS ONE, 2015-08-27;10(8):e0134974.
    Species: Human
    Sample Types: Serum
  10. Neuroblastoma and bone metastases: clinical significance and prognostic value of Dickkopf 1 plasma levels.
    Authors: Granchi D, Corrias MV, Garaventa A, Baglio SR, Cangemi G, Carlini B, Paolucci P, Giunti A, Baldini N
    Bone, 2010-07-14;48(1):152-9.
    Species: Human
    Sample Types: Plasma
  11. Activation of platelets by Aspergillus fumigatus and potential role of platelets in the immunopathogenesis of Aspergillosis.
    Authors: Rodland EK, Ueland T, Pedersen TM, Halvorsen B, Muller F, Aukrust P, Froland SS
    Infect. Immun., 2009-12-14;78(3):1269-75.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Dkk-1 inhibits intestinal epithelial cell migration by attenuating directional polarization of leading edge cells.
    Authors: Koch S, Capaldo CT, Samarin S, Nava P, Neumaier I, Skerra A, Sacks DB, Parkos CA, Nusrat A
    Mol. Biol. Cell, 2009-09-23;20(22):4816-25.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Bone marrow cells are a rich source of growth factors and cytokines: implications for cell therapy trials after myocardial infarction.
    Authors: Korf-Klingebiel M, Kempf T, Sauer T, Brinkmann E, Fischer P, Meyer GP, Ganser A, Drexler H, Wollert KC
    Eur. Heart J., 2008-10-25;29(23):2851-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Paracrine inhibition of osteoblast differentiation induced by neuroblastoma cells.
    Authors: Granchi D, Baglio SR, Amato I, Giunti A, Baldini N
    Int. J. Cancer, 2008-10-01;123(7):1526-35.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Zoledronic acid decreased osteolysis but not bone metastasis in a nude mouse model of canine prostate cancer with mixed bone lesions.
    Authors: Thudi NK, Martin CK, Nadella MV
    Prostate, 2008-07-01;68(10):1116-25.
    Species: Human
    Sample Types: Serum
  16. Clinical, radiographic, and biochemical characterization of multiple myeloma patients with osteonecrosis of the jaw.
    Authors: Raje N, Woo SB, Hande K, Yap JT, Richardson PG, Vallet S, Treister N, Hideshima T, Sheehy N, Chhetri S, Connell B, Xie W, Tai YT, Szot-Barnes A, Tian M, Schlossman RL, Weller E, Munshi NC, Van Den Abbeele AD, Anderson KC
    Clin. Cancer Res., 2008-04-15;14(8):2387-95.
    Species: Human
    Sample Types: Serum
  17. A potential role for Dkk-1 in the pathogenesis of osteosarcoma predicts novel diagnostic and treatment strategies.
    Authors: Lee N, Smolarz AJ, Olson S, David O, Reiser J, Kutner R, Daw NC, Prockop DJ, Horwitz EM, Gregory CA
    Br. J. Cancer, 2007-11-06;97(11):1552-9.
    Species: Human
    Sample Types: Serum
  18. Mesenchymal stem cell abnormalities in patients with multiple myeloma.
    Authors: Garderet L, Mazurier C, Chapel A, Ernou I, Boutin L, Holy X, Gorin NC, Lopez M, Doucet C, Lataillade JJ
    Leuk. Lymphoma, 2007-10-01;48(10):2032-41.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Production of Wnt inhibitors by myeloma cells: potential effects on canonical Wnt pathway in the bone microenvironment.
    Authors: Giuliani N, Morandi F, Tagliaferri S, Lazzaretti M, Donofrio G, Bonomini S, Sala R, Mangoni M, Rizzoli V
    Cancer Res., 2007-08-15;67(16):7665-74.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. The Wnt pathway regulator DKK1 is preferentially expressed in hormone-resistant breast tumours and in some common cancer types.
    Authors: Forget MA, Turcotte S, Beauseigle D, Godin-Ethier J, Pelletier S, Martin J, Tanguay S, Lapointe R
    Br. J. Cancer, 2007-01-23;96(4):646-53.
    Species: Human
    Sample Types: Cell Culture Supernates, Tissue Homogenates

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