Dickkopf-Related Protein 1 Human ELISA

Summary

Dickkopf-1 (Dkk-1) is encoded by the gene dickkopf together with other members of dickkopf protein family in vertebrates, Dkk-2, –3, –4 and a distant family member soggy, sometimes also called Dickkopf-like protein 1 (DKKL1). Dickkopf name is derived from german dick=thick and kopf=head and this protein was independently found and characterised and named as „Sk“. Dkk-1/Sk is composed from 266 amino acids and has predicted molecular weight 25.8kDa. The protein has one N-glycosylation site at the N-terminus. Dkk proteins possess the signal sequence and have two typical cysteine rich domains. The second cysteine-rich domain is required for binding to Lrp6 and Kremen-2. Up to now known receptors are from the family Lrp (Lrp5/6) and Kremen 1 and 2. Dkks are modulating Wnt signalling. Their effect is mostly inhibitory with exception of the Dkk-2 which is activating Wnt-signalling. Wnt signalling pathways are shifting the cell proliferation, cell identity and cell polarity from embryonic to adult homeostasis. Wnt is forming the complex composed of seven-transmembrane receptor Frizzled (Fz) receptor and a lipoprotein-receptor related protein Lrp5 or Lrp6. This triple complex formation is stabilizing the beta-catenin and activating the pathway. Dkk-1 is inhibiting this process by its interaction with Lrp6 that is blocking the Wnt-Fz-Lrp complex formation. The Lrp6-Dkk-1 is entering the cell by endocytosis allowed by the coreceptor Kremen. Dkk-1 in mouse was found to be exprimed in bone, specifically in osteoblasts and osteocytes. The overexpression of the Dkk-1 in Xenopus is causing the ectopic head formation and blocation by anti-Dkk-1 antibodies lead to microcephaly. In mouse models, when missing the Dkk-1, there was found incomplete development of structures anterior to the midbrain and consequently this lead to the perinatal death. In limb development of Dkk-1 null mice there has been found syndactyly and polydactyly. Dkk-1 ELISA is estimating the Dkk-1 in serum or plasma thus can be used in the cancer research, especially bone and lung cancer, as well as in the Paget’s disease or in the problems with bone calcification.

Features

• It is intended for research use only
• The total assay time is less than 3.5 hours
• The kit measures Dkk-1 in serum
• Assay format is 96 wells
• Quality Controls are human serum based
• Standard is recombinant protein based
• Components of the kit are provided ready to use, concentrated or lyophilized

Research topic

Bone and cartilage metabolism, Oncology

Assay format

Sandwich ELISA, Biotin-labelled antibody

Applications

Serum

Sample requirements

singlet 50ul/well

doublet 100ul/well

Storage/Shipping

2 – 8°C

Calibration Curve

Calibration range

0.125 – 4 ng/ml

Limit of detection

0.01 ng/ml

Intra-assay

7,10%

Inter-assay

7,30%

Spiking Recovery

94,50%

Dilution Linearity

93,20%

References to summary

• Adamska M, MacDonald BT, Meisler MH. Hypomorphic expression of Dkk1 in the doubleridge mouse: dose dependence and compensatory interactions with Lrp6. Development. 2004 Jun;131 (11):2543-52
• Davie MW, McCarthy HS, Powell DE, Sharp CA, Evans SF, Marshall MJ. Increased circulating Dickkopf-1 in Paget's disease of bone. Clin Biochem. 2009 Jul;42 (10-11):965-9
• Doriath V, Journe F, Voorzanger-Rousselot N, Body JJ, Garnero P. Assessment of circulating Dickkopf-1 with a new two-site immunoassay in healthy subjects and women with breast cancer and bone metastases. Calcif Tissue Int. 2009 May;84 (5):348-54
• Fedi P, Bafico A, Nieto Soria A, Burgess WH, Miki T, Bottaro DP, Kraus MH, Aaronson SA. Isolation and biochemical characterization of the human Dkk-1 homologue, a novel inhibitor of mammalian Wnt signaling. J Biol Chem. 1999 Jul 2;274 (27):19465-72
• Karsenty G, Glass DA 2nd. In vivo analysis of Wnt signaling in bone. Endocrinology. 2007 Jun;148 (6):2630-4
• Lee N. Smolarz AJ. Olson S. David O. Reiser J. Kutner R. Daw NC. Prockop DJ. Horwitz EM and Gregory CA. A potential role od Dkk-I in the pathogenesis of osteosarcoma predicts novel diagnostic and treatment strategies. British Journal of Cancer. 97:1552-1559 (2007);
• Marshall MJ. Evans SF. Sharp CA. Powell DE. McCarthy HS. Davie MWJ. Increased circulating Dickkopf-1 in Paget´s disease of bone. Clin Biochem. 42: 965-969 (2009);
• Niehrs C. Function and biological roles of the Dickkopf family of Wnt modulators. Oncogene. 2006 Dec 4;25 (57):7469-81
• Qin WX, Yu B, Huang G, Sheng SL. Clinical significance and prognostic value of serum Dickkopf-1 concentrations in patients with lung cancer. Clin Chem. 2009 Sep;55 (9):1656-64
• Sheng SL, Huang G, Yu B, Qin WX. Clinical significance and prognostic value of serum Dickkopf-1 concentrations in patients with lung cancer. Clin Chem. 2009 Sep;55 (9):1656-64
• Yamabuki T, Takano A, Hayama S, Ishikawa N, Kato T, Miyamoto M, Ito T, Ito H, Miyagi Y, Nakayama H, Fujita M, Hosokawa M, Tsuchiya E, Kohno N, Kondo S, Nakamura Y, Daigo Y. Dickkopf-1 a Novel Serologic and Prognostic Biomarker for Lung and Esophageal carcinomas. Cancer Res. (2007);

By research topic:

• Bone and cartilage metabolism
• Oncology

By molecule:

• Dickkopf-Related Protein 1

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