Human dkk1 quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States

The Human DKK1 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human Dickkopf-related protein 1 (DKK1) levels in cell culture supernatants, serum, and plasma. It utilizes a microplate pre-coated with a monoclonal antibody specific for DKK1.

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Lab products found in correlation

Mouse dkk 1 quantikine elisa kit, by R&D Systems (2 mentions) Xmark microplate absorbance spectrophotometer, by Bio-Rad (1 mentions) Human mcp 1 quantikine elisa kit, by R&D Systems (1 mentions) Spectrostar, by BMG Labtech (1 mentions) Af1096, by R&D Systems (1 mentions) Eon microplate spectrophotometer, by Agilent Technologies (1 mentions) Anti dkk1, by R&D Systems (1 mentions) Liproxstatin 1, by Targetmol (1 mentions) Xav 939, by Targetmol (1 mentions) Chir99021, by Targetmol (1 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Aldefluor kit, by STEMCELL (1 mentions) Chip assay kit, by Beyotime (1 mentions) Erastin, by MedChemExpress (1 mentions) Autokit micro albumin, by Fujifilm (1 mentions)

12 protocols using human dkk1 quantikine elisa kit

Dkk-1 Quantification in MSC Secretome

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Protein levels of Dkk-1 were determined using Human Dkk-1 Quantikine ELISA Kit (R&D Systems™). A serum-free conditioned culture medium obtained from 1 × 10⁵ MSC (unexposed or exposed to DOX or PAC for 48 h) was used in the assay according to the manufacturer´s protocol. Absorbance was measured on the xMark™ Microplate Absorbance Spectrophotometer (Bio-Rad Laboratories Ltd, Watford, UK).

Plava J., Burikova M., Cihova M., Trnkova L., Smolkova B., Babal P., Krivosikova L., Janega P., Rojikova L., Drahosova S., Bohac M., Danisovic L., Kucerova L, & Miklikova S. (2021). Chemotherapy-triggered changes in stromal compartment drive tumor invasiveness and progression of breast cancer. Journal of Experimental & Clinical Cancer Research : CR, 40, 302.

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Quantification of Secreted DKK1

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Concentrations of secreted DKK1 in culture supernatant were determined using a Human DKK1 Quantikine ELISA Kit (R&D systems, DKK100) according to the manufacturer’s protocols. Briefly, after lentivirus-mediated infections with shG9a- or G9a/V5-expressing vector and proper selections with antibiotics, equal numbers of infected cells were re-plated in 96-well plates. After 72 h of additional culture, the culture supernatants were harvested and subsequently subjected to ELISA. Also, culture supernatants were harvested from DMSO- and UNC0638-treated cells 72 h after treatment. All of the supernatant samples were stored at −80°C after removal of cell debris by centrifugation.

Kato S., Weng Q.Y., Insco M.L., Chen K.Y., Muralidhar S., Poźniak J., Diaz J.M., Drier Y., Nguyen N., Lo J.A., van Rooijen E., Kemeny L.V., Zhan Y., Feng Y., Silkworth W., Powell C.T., Liau B.B., Xiong Y., Jin J., Newton-Bishop J., Zon L.I., Bernstein B.E, & Fisher D.E. (2020). Gain-of-function genetic alterations of G9a drive oncogenesis. Cancer discovery, 10(7), 980-997.

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Serum DKK-1 Levels in Hepatocellular Carcinoma

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Serum DKK-1 levels were measured by ELISA using stored sera from 400 patients with HCC and 205 patients with CLD without HCC as control at the Kanazawa University Hospital from October 2002 to October 2017. Surgically resected HCC samples previously used [41 (link)] were evaluated for the expression of EpCAM and AFP as previously described [18 (link)]. We included 14 and 44 HCC samples of HpSC-HCC and MH-HCC, respectively, in the study. Among them, RNA samples and sera were available for 8 HpSC-HCC and 35 MH-HCC or 14 HpSC-HCC and 36 MH-HCC specimens, respectively. The Human Dkk-1 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) was used for the measurement of serum DKK-1 levels. The final value of the DKK-1 level was obtained after correction using a standard curve. The clinical information of the patients was collected retrospectively from medical records. The study conformed to the standards set by the Declaration of Helsinki, and the protocol was approved by the institutional review board of the Graduate School of Medical Sciences, Kanazawa University (IRB number: 2016-093).

Suda T., Yamashita T., Sunagozaka H., Okada H., Nio K., Sakai Y., Yamashita T., Mizukoshi E., Honda M, & Kaneko S. (2022). Dickkopf-1 Promotes Angiogenesis and is a Biomarker for Hepatic Stem Cell-like Hepatocellular Carcinoma. International Journal of Molecular Sciences, 23(5), 2801.

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Quantification of DKK-1, IL-6, and TGF-β1 in DPC-conditioned Medium

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For the quantification of DKK-1 in DPC-conditioned medium, the Human DKK-1 Quantikine ELISA Kit (R&D systems, Minneapolis, MN, USA, #DKK100B) was used, according to the supplier’s protocol. DPCs were cultured to 80% confluency and then treated with DHT. Conditioned media was acquired and preserved in − 80 °C. After centrifugation at 3000 g for 5 min, supernatants were used for the ELISA assay. IL-6 and TGF-β1 concentrations in DPC-conditioned medium were measured with human IL-6 (LabisKOMA, Seoul, Korea, #K0331194) and TGF-β1 (LabisKOMA, Seoul, Korea, #K0332110) ELISA kits.

Jung Y.H., Chae C.W., Choi G.E., Shin H.C., Lim J.R., Chang H.S., Park J., Cho J.H., Park M.R., Lee H.J, & Han H.J. (2022). Cyanidin 3-O-arabinoside suppresses DHT-induced dermal papilla cell senescence by modulating p38-dependent ER-mitochondria contacts. Journal of Biomedical Science, 29, 17.

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Quantification of Serum DKK1 by ELISA

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The serum concentration of DKK1 was measured by Human DKK1 Quantikine ELISA Kit (R & D system) according to the manufacturer's instructions. Briefly, 100 μL Assay Diluent was added into each well, followed by 100 μL standard, control or sample and the microplate was incubated at room temperature for 2 h. Then each well was washed 3 times, blocked with 200 μL Conjugate and incubated for 2 h at room temperature, followed by another washing step. In addition, 200 μL Substrate Solution was added and incubated at 37°C for 30 mins without light. Finally, the reaction was terminated by adding 50 μL Stop Solution. The absorbance was read at a 450 nm wavelength. The concentration of DKK1 was evaluated based on standard curves.

Jia X., Li N., Peng C., Deng Y., Wang J., Deng M., Lu M., Yin J., Zheng G., Liu H, & He Z. (2016). miR-493 mediated DKK1 down-regulation confers proliferation, invasion and chemo-resistance in gastric cancer cells. Oncotarget, 7(6), 7044-7054.

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Quantifying DKK1 and MCP-1 in Plasma

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The plasma samples were diluted four times with Calibrator Diluent and DKK1 amount measured in duplicates using the Human DKK1 Quantikine ELISA Kit (DKK100), according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN). Bound DKK1 signal was developed with 100μl of tetramethylbenzemidine microwell peroxidase substrate system (KPL, #50-76-00). The reaction was stopped with addition of 100μl 1M H₃PO₄. The optical density was acquired in a microplate reader set to 450 nm and a correct set to 540 nm. DKK1 levels were calculated from a standard curve generated from DKK1 standard in the same plate. Human plasma MCP-1 levels were similarly measured using Human MCP-1 Quantikine ELISA Kit (DCP00, R&D Systems) per manufacturer’s instruction.

YU C., SEATON M., LETENDRE S., HEATON R, & AL-HARTHI L. (2017). Plasma Dickkopf-related Protein 1, an antagonist of the Wnt pathway, is associated with HIV-associated neurocognitive impairment. AIDS (London, England), 31(10), 1379-1385.

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Quantifying Serum DKK1 Levels in Periodontal Disease

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DKK1 serum levels at baseline and after periodontal therapy were measured using an enzyme-linked immunosorbent assay (ELISA) (Human DKK1 Quantikine ELISA Kit; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol. The microplate was precoated with human DKK1 monoclonal antibody. Standard or diluted serums (8-fold dilution) were added per well and incubated for 2 hours. After washing, human DKK1 conjugate was added to each well, incubated for 2 hours, and then washed. Substrate was added for 30 minutes, and then the reaction was stopped. The optical density of each well was determined at 450 nm using an ELISA spectrophotometric reader (SPECTROstar; BMG LABTECH, Offenburg, Germany). The total amounts of DKK1 were displayed as picogram/milliliter (pg/mL).

Azab E., Attia A., Yaghmoor W., Aldahlawi S, & Youssef A.R. (2022). The Impact of Nonsurgical Periodontal Therapy on Serum Levels of Dickkopf-Related Protein-1 in Smokers and Nonsmokers with Periodontitis: A Prospective Comparative Study. Clinical, Cosmetic and Investigational Dentistry, 14, 191-198.

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Quantitative DKK1 Measurement Protocols

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Secreted DKK1 levels were measured either by Western blot after immunoprecipitating DKK1 from the culture medium using anti-DKK1 (#AF1096; R&D Systems) or by commercially available ELISA assay (Human Dkk-1 Quantikine ELISA Kit, #DKK100B; R&D Systems) according to the manufacturer’s protocol. For DKK1 ELISA, absorbance was measured at 450 nm with a correction at 540 nm using the Eon microplate spectrophotometer (Agilent).

Kim H., Jang S, & Lee Y.S. (2022). The m6A(m)-independent role of FTO in regulating WNT signaling pathways. Life Science Alliance, 5(5), e202101250.

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Quantification of Serum DKK1 Levels

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Serum or plasma levels of DKK1 from mice or patients were quantified using ELISA kits specific for DKK1 (Mouse Dkk-1 Quantikine ELISA kit or Human Dkk-1 Quantikine ELISA kit, R&D systems) as per the manufacturer’s protocols.

Faccio R., Lee S., Ricci B., Tran J., Ye J., Clever D., Eul E., Wang J., Wong P., Ma C, & Fehniger T. (2024). Cancer-associated fibroblast-derived Dickkopf-1 suppresses NK cell cytotoxicity in breast cancer. Research Square.

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Quantifying Maternal DKK1 Levels

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Maternal secreted levels of DKK1 were measured using a sandwich enzyme-Linked immunosorbent Assay (ELISA) kit (Human DKK1 Quantikine ELISA Kit, catalog no. DKK100B, R & D Systems, Germany). Measurement procedures were done following the manufacturerʼs instruction manual. All measurements were performed in duplicate. Optical density was measured at 450 nm and referenced to 570 nm on a 96-well microplate reader (Sunrise-Tecan, Life Science) and protein levels were obtained with a four-parameter logistic curve fitted against a standard curve and multiplied by the dilution factor using Magellan 7.2 Ink Data Analysis Software (Life Science-Tecan).

Kasoha M., Takacs Z., Fackiner L., Gerlinger C., Sklavounos P., Radosa J., Solomayer E.F, & Hamza A. (2021). Comparison of Maternal Serum Levels and Placental mRNA Levels of Dickkopf-1 in Preeclamptic and Normal Pregnant Women at Delivery. Geburtshilfe und Frauenheilkunde, 81(11), 1247-1255.

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