Lbis mouse albumin elisa kit

The unbound fraction was determined by equilibrium dialysis using a rapid equilibrium dialysis device (Thermo Fisher Scientific, Waltham, MA, USA). Paclitaxel or [³H(G)]digoxin ([³H] digoxin, 39.8 Ci/mmol; PerkinElmer, Boston, MA, USA) was spiked into maternal or fetal plasma at a final concentration of 26 or 13 nM. Spiked plasma (100 μL) was loaded into the sample chambers of the device, and the buffer chambers were filled with 350 μL of phosphate-buffered saline. The device was covered with sealing tape and incubated for 12 h at 37°C on an orbital shaker running at 250 rpm. Following incubation, aliquots from both chambers were taken for measurement of Paclitaxel concentration or radioactivity of [³H]digoxin using LC-MS/MS or liquid scintillation counting, respectively. The unbound fraction in the maternal and fetal plasma (f_u,mp and f_u,fp) was calculated as the PBS-to-plasma concentration (radioactivity) ratio. Albumin concentration in maternal and fetal plasma was determined using an LBIS Mouse Albumin ELISA Kit (Fujifilm Wako Shibayagi, Shibukawa, Japan) according to the manufacturer’s protocol.

The level of blood glucose was determined using tail blood and a OneTouch UltraVue (Johnson & Johnson) or Antsense III (Horiba) glucometer. Urinary albumin was measured in the 24-h urine collections using an LBIS Mouse Albumin ELISA Kit according to the manufacturer's instructions (Fujifilm). The urine creatinine concentration of the 24-h urine was detected by using the LC‒MS/MS method, and urine osmolality was examined by a freezing point depression method. For assessment of fractional excretion of glucose, the urine glucose concentration was measured by using a Glucose Assay Kit-WST according to the manufacturer's instructions (Dojindo).

BMDMs were incubated in culture plates and treated with 1 μg/ml LPS either alone or with 2.5% (v/v) concentration of JP-treated serum. The concentrations of TNF-α and interleukin (IL)-6 in the supernatant of BMDMs were measured with enzyme-linked immunosorbent assay (ELISA) kits (NOVUS biologicals, CO, USA).
The urine of the mice was centrifuged at 12,000 r/m for 15 min to collect the supernatant, and then the urine protein concentration in the urine was measured according to the LBIS Mouse Albumin ELISA Kit (FUJIFILM, Gunma, Japan). Each mouse venous blood was allowed to stand for 30 min, and the serum was collected by centrifugation. Afterwards, the concentration of anti-dsDNA in the serum was measured with the LBIS Mouse anti-dsDNA ELISA Kit (FUJIFILM, Gunma, Japan). The procedure was strictly in accordance with the kit instructions and the required indicators were measured in a microplate reader (PerkinElmer, EnSpire, MA, United States).