BHPrS-1 or NHPrE-1 cells were seeded in 6-well plates at 100,000 cells/well density. Treatments IL-1β (1, 10 or 100 pg) or TGF-β1 (0.1, 1 or 10 ng) were added in RPMI supplemented with 1% heat-inactivated FBS for BHPrS-1 and DMEM/F12 with 0.1% BSA for NHPrE-1 cells. Media were collected 24 hours later and cell debris was removed by centrifugation at 3000 rpm for 3 min and stored at −80°C. The concentration of OPN was determined by the Human Osteopontin (OPN) DuoSet ELISA (DY1433, R&D Systems) according to the manufacturer`s instructions. OPN concentration was calculated by GraphPad Prism ver. 8 using second order polynomial regression analysis.
Opn duoset elisa
Manufactured by R&D Systems
Sourced in United States
The R&D Systems OPN DuoSet ELISA is a sandwich enzyme-linked immunosorbent assay (ELISA) kit designed for the quantitative measurement of an analyte of interest in cell culture supernates, serum, and plasma samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using opn duoset elisa
1
Osteopontin Secretion in Prostate Cells
2
Growth Factor Release from Hydrogels
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The release of bone-related growth factors from HPS-F and NS-F was evaluated over 7 days. The growth factors examined included vascular endothelial growth factor-A (VEGF-A), OPG, OPN, platelet-derived growth factor BB (PDGF-BB), basic fibroblast growth factor (bFGF), and sRANKL. For the experimental setup, HPS-F and NS-F were prepared as described in Section 4.3 for 1 mL of HPS and NS, respectively, and transferred into a 5 mL tube (Eppendorf, Hamburg, Germany). After a gelation time of 1 min, 1 mL of PBS was added, and the tubes were incubated on a shaker plate at room temperature. The supernatants were collected after 12 h and after 1, 2, 3, 4, 5, 6, and 7 days. Samples were stored at −80 °C until ELISA measurements were taken. ELISAs were carried out according to the manufacturing protocol (VEGF DuoSet ELISA (Catalog # DY293B); OPG DuoSet ELISA (Catalog # DY805); OPN DuoSet ELISA (Catalog #: DY1433) or PDGF-BB DuoSet ELISA (Catalog #: DY220); bFGF DuoSet ELISA (Catalog #: DY233) R&D Systems, Minneapolis, MN, USA). Readout was performed through measurement of the optical density using a Mithras LB 940 Multimode Microplate Reader (Berthold Technologies GmbH & Co. KG, Bad Wildbad, Germany).
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