Human osteopontin elisa kit

Protein levels of markers for osteoblastic differentiation (c-terminal pro-peptide of collagen type 1 (C1CP): MicroVue™ CICP EIA, Quidel, Marburg, Germany; osteopontin (OPN): Human Osteopontin ELISA Kit, Abcam, Cambridge, UK), IL-6 signaling (human sgp 130 ELISA, Sigma-Aldrich, ST. Louis MO, USA; IL-6: Human IL-6 ELISA Ready-SET-Go!®, ThermoFisher Scientific, Waltham, MA, USA; sIL-6R: IL-6 Receptor Human ELISA Kit, Abcam, Cambridge, UK) as well as IL-8 (Human IL-8 ELISA Ready-SET-Go!®, ThermoFisher Scientific, Waltham, MA, USA) were determined in the supernatant of control and exposed osteoblasts via enzyme-linked immunosorbent assay. For this purpose, supernatants were collected and stored at -20 °C before quantification. ELISAs were performed according to the manufacturer's recommendations. Absorbance was measured at 405 nm (reference wavelength: 630 nm; IL-6: 570 nm) using the Tecan Infinite® 200 Pro microplate reader (Tecan Group AG, Maennedorf, Switzerland). The sample concentrations were calculated using a standard curve. Normalization of protein content to total protein was performed using the Qubit Protein Assay Kit and Qubit 1.0 (both: Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions.

We performed enzyme-linked immunosorbent assays (ELISAs) for osteopontin, MMP-7, neuropilin-1, or prostate specific antigen (PSA), following manufacturer’s instructions. We used the following kits: Human Osteopontin ELISA Kit (Abcam, Cambridge, UK, #ab192143), Human Neuropiln-1 ELISA Kit (Abcam #ab227901), Human Total Prostate-Specific Antigen ELISA Kit (Abcam #ab188388), and Quantikine ELISA Human Total MMP-7 (R&D Systems Minneapolis, MN, #DMP700). Plates were read in a Tecan Infinite 200 Pro M Nano plate reader at 450nm.