Human tgf β elisa kit

CXCL13 levels in conditioned medium, BM, and peripheral blood plasma from MM patients and healthy volunteers were examined using a human CXCL13 ELISA kit (RayBiotech) according to the manufacturer's instructions. TGFβ levels in conditioned medium were examined using a human TGFβ ELISA kit (R&D Systems) according to the manufacturer's instructions. The murine RANKL levels in the plasma of non-inoculated (n = 3), RPMI8226-CXCR4-GFP-inoculated (n = 4) or RPMI8226-CXCR4-GFP-CRISPR-CXCL13-inoculated (n = 4) animals were determined using murine RANKL ELISA kit (R&D systems) according to the manufacturer's instructions.

The concentrations of IL-6 and IL-10 were determined by cytometric bead array (CBA) using a commercial kit (BD Biosciences, San Joes, USA), and were quantified using the Cell Quest Pro and CBA software (Becton–Dickinson) on a FACS Calibur cytometry. The concentrations of transforming growth factor-β (TGF-β) in individual subjects were determined by enzyme linked immunosorbent assay (ELISA) using a human TGF-β ELISA kit (R&D Systems, Minneapolis, USA) and were calculated, according to the standard curve established using the recombinant TGF-β provided. The limitation of detection for IL-6, IL-10 or TGF-β was 2.5, 3.3, or 15.4 pg/mL, respectively.

Keloid-derived fibroblasts were incubated with LY2109761 (5 or 10 μM) or vehicle control (DMSO) for 48 h. After treatment, the supernatant was collected and the concentration of TGF-β was measured using a human TGF-β ELISA kit (R&D, San Jose, CA, USA). Absorbance was measured at 450 nm using a microreader (Thermo Fisher Scientific, Waltham, MA, USA). The assay was performed in triplicate and repeated in three cell samples.