Human osteopontin quantikine elisa kit

Pharmacodynamic assays were performed to assess the activity of ASP5094 on the known downstream effectors of α9 in RA of TNF-α, MMP-3, and IL-6, which were measured at baseline and each scheduled visit. These serum concentrations were measured by commercially available kits as follows: TNF-α; ELISA, Quantikine HS ELISA (R&D Systems), MMP-3; LTIA, Panaclear MMP-3: Latex (Sekisui Medical), IL-6; CLEIA, IL-6 LPG Immunoreaction Cartridges (Fujirebio). Levels of putative α9 ligands tenascin (TNC)-C (large variant of tenascin-C, termed FNIII-B variant), vascular cell adhesion molecule (VCAM)-1, and osteopontin (OPN; full-length and thrombin-cleaved) were determined at baseline and week 12/discontinuation and week 16/discontinuation, then 4 weeks later for exploratory evaluation of ASP5094 on potential biomarkers. Plasma concentrations of TNC-C, VCAM-1, and OPN were measured by a commercially available ELISA kit as follows: TNC-C; Tn-C large [FNIII-B] assay kit (Immuno-Biological Laboratories), VCAM-1; Human sVCAM1/CD106 Quantikine ELISA kit; (R&D Systems), OPN full-length; Human osteopontin Quantikine ELISA kit (R&D Systems), cleaved-OPN; Human osteopontin N-half assay kit (Immuno-Biological Laboratories).

RPMI-1640, FBS, and Pencillin/Streptomycin/Amphotercin B (PSA) were purchased from CellGro; IMDM medium was purchased from Thermo Scientific; OptiMEM was purchased from Invitrogen; Puromycin (Sigma-Aldrich, Inc.); Quantikine human osteopontin ELISA kit (R & D Systems, Inc.); Rabbit anti-osteopontin (Rockland Immunochemicals, Inc. Limerick, PA); molecular biology-grade reagents (Sigma); Trizol and PureLink RNA extraction kit (Life Technologies); Nano 6000 RNA analysis reagents (Agilent); PCR primers and TaqMan FAST Universal PCR Master Mix (Life Technologies); BD Biocoat Matrigel Invasion Chamber (BD Biosciences); Recombinant OPN (R & D Systems, Inc.).