Human DKK-1 ELISA Assay Kit

$590.00

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

SKU: BI-20413 Categories: , ,

Human DKK-1 ELISA Assay Kit

For Research Use Only

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
Controls Included

Product Developed and Manufactured by Biomedica


Unit conversion: 1 pg/ml = 0.039 pmol/l (MW = 25.8 kDa)

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

 

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.

Manual

Product Manual


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.