Mouse fgf23 elisa kit

Active FGF23 and DMP1 intracellular and extracellular levels were evaluated in cell lysates and culture media using mouse FGF23 ELISA kit and DMP1 ELISA Kit (Abcam, Cambridge, UK), respectively, according to the manufacturer’s instructions. MLO-Y4 cells were seeded in 12-well plates and treated with 17β–E2 or inhibitors for 4, 24 or 48 h, as described above. Cell lysates were performed detaching and collecting MLO-Y4 by centrifugation at 130× g for 10 min. Cells, washed in cold phosphate buffer saline (PBS), were suspended in PBS and sonicated four times. Then, cell lysates were centrifuged at 1500× g for 10 min at 4 °C and FGF23 and DMP1 assays were performed in the supernatants. Absorbance was measured at 450 nm with iMarkTM Microplate Absorbance Reader (BioRad, Hercules, CA, USA). Data were normalized on total protein content and FGF23 and DMP1 levels were expressed as a percentage of the respective levels measured in control set as 100%.

A commercially available clinical chemistry analyzer (Roche, Rotkreuz, Switzerland) was used to measure the serum albumin, globulin, creatinine, and blood urea nitrogen (BUN) levels. A mouse ELISA kit (Immunology Consultants Laboratory, Portland, OR, USA) was used to detect the levels of cystatin C. Serum fibroblast growth factor (FGF)-23 was determined using a mouse FGF23 ELISA kit (Abcam, Cambridge, MA, USA).

Serum FGF-23 was quantified using mouse FGF-23 ELISA kit (Abcam, Waltham, MA). The kit components were reconstituted, and the assay was performed according to the manufacturer’s instruction. Data are presented as picogram/milliliter.