Levels of human CD14 in serum samples from 19 HCC patients and 18 LC patients were assayed using a commercial ELISA kit (Human sCD14 Quantikine ELISA, R&D, USA) according to the manufacturer’s instructions. Experiments were performed in duplicate.
Human scd14 quantikine elisa
Manufactured by R&D Systems
Sourced in United States, Switzerland, China
The Human sCD14 Quantikine ELISA is a quantitative sandwich enzyme immunoassay designed to measure soluble CD14 levels in cell culture supernates, serum, and plasma. It is a tool for researchers to quantify this specific analyte in their samples.
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4 protocols using human scd14 quantikine elisa
1
Serum CD14 Levels in HCC and LC
2
Quantification of Plasma sCD14 by ELISA
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Plasma sCD14 levels were determined using the Human sCD14 Quantikine® ELISA (R&D Systems). Plasma samples were diluted 200-fold with sample diluent solution prior to analysis. Optical density was determined using a Microplate Reader 3550 (BioRad) and plasma sCD14 concentrations were calculated using the Microplate Manager 4.0 (BioRad).
3
Plasma LPS, sCD14, LBP, and D-Dimer Assays
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Limulus Amebocyte Lysate (LAL) (Lonza; Basel, Switzerland) and Human sCD14 Quantikine ELISA (R&D, Minneapolis, Minnesota, USA) were used to measure plasma lipopolysaccharide (LPS) and soluble CD14 (sCD14), as described [8, 25] LPS binding protein (LBP) (Hycult Biotech, Uden, The Netherlands), and D-Dimer (Technoclone, Vienna, Austria) were determined in plasma samples by ELISA according to the manufacturers' instructions.
4
Maternal and Newborn CRP and sCD14 Levels
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Non-fasting blood samples were collected in serum clotting tubes from mothers at approximately 28 weeks gestation. Umbilical cord blood (≤ 5ml) was collected before delivery of the placenta from placental vessels into a serum clotting tube.
Both maternal and newborn blood samples were centrifuged within 2 hours of collection and the serum aliquoted and stored at -80 0 C before assaying.
High-sensitivity CRP was measured on a COBAS Integra 400 plus Analyser (Roche Diagnostics Australia, Castle Hill, NSW, Australia); maximum recorded value was 20mg/dL. Soluble CD14 serum levels were assayed using the Human sCD14 Quantikine ELISA (R&D Systems, China).
Both maternal and newborn blood samples were centrifuged within 2 hours of collection and the serum aliquoted and stored at -80 0 C before assaying.
High-sensitivity CRP was measured on a COBAS Integra 400 plus Analyser (Roche Diagnostics Australia, Castle Hill, NSW, Australia); maximum recorded value was 20mg/dL. Soluble CD14 serum levels were assayed using the Human sCD14 Quantikine ELISA (R&D Systems, China).
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