DKK-1 ELISA Assay Kit

$610.00

The DKK-1 ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of Human DKK-1 in serum. The Eagle Biosciences DKK-1 ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

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DKK-1 ELISA Assay Kit

DKK-1 ELISA Assay Kit Developed and Manufactured in Austria by Biomedica

Size: 1×96 wells
Sensitivity: LOD: 1.7 pmol/l (0 pmol/l + 3 SD) , LLOQ: 1.25 pmol/l
Dynamic Range: 0 to 160 pmol/l
Incubation Time: 3.5 hours
Sample Type: Human serum
Sample Size: 20 µL
Alternative Names: DKK-1 ELISA, DKK1 ELISA, Dickkopf-1 ELISA, Human Dickkopf-1 ELISA
For Research Use Only
Controls Included


Unit conversion: 1 pg/ml = 0.039 pmol/l (MW = 25.8 kDa)
Reference Range: Median Serum (n=51)= 34 pmol/l
Each laboratory should establish its own reference range for the samples under investigation.
Specificity: oligomeric forms of natural and recombinant human DKK-1


Assay Principle

The DKK-1 ELISA Assay kit is a sandwich enzyme immunoassay for the quantitative determination of DKK-1 in human serum samples. In a first step, assay buffer, standard/control/sample and detection antibody (biotinylated polyclonal rabbit anti-human DKK-1) are pipetted into the wells of the microtiter strips, which are pre-coated with monoclonal mouse anti-human DKK-1 antibody. DKK-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step all non-specific unbound material is removed. In a next step, the conjugate (streptavidin-HRP) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of DKK-1 present in the sample. This color change is detectable with a standard microtiter plate reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated using the values obtained from the standard. The concentration of DKK-1 in the sample is determined directly from the dose response curve.


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Additional Information

Assay Background


Dickkopf-1(DKK-1) is a 28.67 kDa secreted protein that acts as soluble inhibitor of the WNT signalling pathway. This pathway contains lipid-modified glycoproteins that activate cell surface receptor-mediated signal transduction to regulate cell activities like: cell fate, proliferation, migration, polarity and gene expression. DKK-1 regulates developmental processes of all kinds. Thus, DKK-1 is also involved in the regulation of bone metabolism as it effects osteoblast differentiation and in regulation of tumourgenetic activity.

Assay Procedure


  1. Pipette 50 µl ASYBUF (assay buffer) into all wells.
  2. Add 20 µl STD/CTRL/SAMPLE (standards/control/sample) in duplicate into respective wells, swirl gently.
  3. Add 50 µl AB (biotinylated DKK-1 antibody) into each well, swirl gently.
  4. Cover tightly and incubate for 2 hours at room temperature (18-24°C).
  5. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (wash buffer). Remove remaining WASHBUF by hitting plate against paper towel after the last wash.
  6. Add 100 µl CONJ (conjugate) into each well.
  7. Cover tightly and incubate for 1 hour at room temperature (18-24°C).
  8. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (wash buffer). Remove remaining WASHBUF by hitting plate against paper towel after the last wash.
  9. Add 100 µl SUB (substrate) into each well.
  10. Incubate for 30 min at room temperature (18-24°C) in the dark.
  11. Add 50 μl STOP (stop solution) into each well, swirl gently.
  12. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

human-dkk-1-elisa-assay-kit

Typical Standard Curve


Human DKK-1 Standard Curve

Expected Values


Values from apparently healthy individuals:

  • Median Serum (n=51)= 34 pmol/l

It is recommended to establish the normal range for each laboratory.

Documents

Product Documents


Publications

Citations


  • Circulating Dickkopf-1 and Radiological Progression in Patients with Early Rheumatoid Arthritis Treated with Etanercept. Garnero P, Tabassi NC, Voorzanger-Rousselot N. J Rheumatol, 2008; 35(12):2313-5.
  • Serum concentrations of DKK-1 correlate with the extent of bone disease in patients with multiple myeloma. Kaiser M, Mieth M, Liebisch P, Oberlander R, Rademacher J, Jakob C, Kleeberg L, Fleissner C, Braendle E, Peters M, Stover D, Sezer O, Heider U. Eur J Haematol, 2008; 80(6):490-4.
  • Low circulating Dickkopf-1 and its link with severity of spinal involvement in diffuse idiopathic skeletal hyperostosis. Senolt L, Hulejova H, Krystufkova O, Forejtova S, Cerezo LA, Gatterova J, Pavelka K, Vencovsky J. Ann Rheum Dis, 2012; 71(1):71-4.
  • Biomarkers of Bone Metabolism in Ankylosing Spondylitis in Relation to Osteoproliferation and Osteoporosis. Klingberg E, Nurkkala M, Carlsten H, Forsblad-d’Elia H. J Rheumatol, 2014; 41: 1349-1356.
  • Bone microstructural changes revealed by high-resolution peripheral quantitative computed tomography imaging and elevated DKK1 and MIP-1α levels in patients with MGUS. Ng AC, Khosla S, Charatcharoenwitthaya N, Kumar SK, Achenbach SJ, Holets MF, McCready LK, Melton LJ, Kyle RA, Rajkumar SV, Drake MT. Blood, 2011; 118: 6529-6534.
  • High dickkopf-1 levels in sera and leukocytes from children with 21-hydroxylase deficiency on chronic glucocorticoid treatment. Brunetti G, Faienza MF, Piacente L, Ventura A, Oranger A, Carbone C, Di Benedetto A, Colaianni G, Gigante M, Mori G, Gesualdo L, Colucci S, Cavallo L, Grano M. Am J Physiol Endocrinol Metab, 2013; 304: E546-E554.
  • DKK-1 in prostate cancer diagnosis and follow up. D’Amelio P, Roato I, Oderda M, Soria F, Zitella A, Ferracini R, Mengozzi G, Gontero P, Isaia GC. BMC Clinical Pathology, 2014; 14:11.
  • Dickkopf-1 as a mediator and novel target in malignant bone disease. Rachner TD, Göbel A, Benad-Mehner P, Hofbauer LC, Rauner M. Cancer Letters, 2014; 346:172–177.