Mouse Dkk-1 Quantikine ELISA Kit

  (4 citations)
(1 Review)
    
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Product Details
Assay Procedure
Citations (4)
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  • 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 (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    29.6 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse Dkk-1.  This assay cross-reacts 94% with recombinant Rat Dkk-1 and 26% with recombinant Human Dkk-1.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse Dkk-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure Dkk-1 in cell culture supernates, mouse serum, and plasma. It contains NS0-expressed recombinant mouse Dkk-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse Dkk-1. Results obtained using natural mouse Dkk-1 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse Dkk-1.

Precision
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
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 230 478 1487 219 481 1492
Standard Deviation 20.1 29.7 80.8 20.8 44.7 144
CV% 8.7 6.2 5.4 9.5 9.3 9.7

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 99 88-107
EDTA Plasma (n=4) 109 93-117
Heparin Plasma (n=4) 113 104-119
Serum (n=4) 113 100-120
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse Dkk-1 in each matrix were diluted with Calibrator Diluent and assayed.
 Dkk-1 [HRP]
Product Datasheets

Certificate of Analysis Lookup

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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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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.

4 Citations: Showing 1 - 4
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Species
Sample Type
  1. Leukocyte Beta-Catenin Expression Is Disturbed in Systemic Lupus Erythematosus
    PLoS ONE, 2016;11(8):e0161682.
    Species: Mouse
    Sample Type: Serum
  2. Dickkopf-1 promotes hematopoietic regeneration via direct and niche-mediated mechanisms
    Nat. Med, 2016;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  3. Bone marrow stromal cells create a permissive microenvironment for myeloma development: a new stromal role for Wnt inhibitor Dkk1.
    Authors: Fowler J, Mundy G, Lwin S, Edwards C
    Cancer Res, 2012;72(9):2183-9.
    Species: Mouse
    Sample Type: Serum
  4. Prostaglandin E(2) Signals Through PTGER2 to Regulate Sclerostin Expression.
    Authors: Genetos DC, Yellowley CE, Loots GG
    PLoS ONE, 2011;6(3):e17772.
    Species: Mouse
    Sample Type: Cell Lysates

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Average Rating: 5 (Based on 1 review)

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