Fuji dri chem 3500v

Blood samples were allowed to clot and were centrifuged to prepare serum. The concentration of total hsFLT1 were measured with ELISA kits according to the manufacturer’s instructions (R&D Systems). Urine albumin and creatinine concentrations were measured using Fuji DRI-CHEM 3500 V and DRICHEM slides (Fujifilm Co, Ltd, Tokyo, Japan) (Skylight Biotech, Inc, Akita, Japan).

On the day of necropsy, blood samples were obtained from the vena cava under isoflurane anesthesia. For clinical chemistry analyses, all blood samples were centrifuged at 1,500 × g for 15 min at 4°C, after which the serum was collected. β-OHB was measured using Precision Xceed (Abbott Japan Co., Ltd., Tokyo, Japan). A hematological analyzer XT-1800iV (SYSMEX Corp., Hyogo, Japan) and a clinical chemistry analyzer (Fuji Drichem 3500V, FUJIFILM Medical Co. Ltd., Tokyo, Japan) were used for hematological assessment and serum chemistry respectively, according to manufacturer’s instructions.

The serum alanine aminotransferase (ALT) levels of the immunized mice were measured using an automatic analyzer (Fuji DRI-chem 3500V; FUJIFILM, Tokyo, Japan). The levels of anti-OVA IgG antibodies in the sera were determined using an enzyme-linked immunosorbent assay (ELISA), as previously described (34 (link)). In brief, ELISA plates were coated with 100 μg/well of the OVA protein (Wako) in PBS overnight at 4°C. The coated plates were washed with PBS containing 0.05% Tween 20 (washing buffer) and blocked with PBS containing 10% FBS for 30 min at room temperature. After the wash, serum (×40) was added to the plates, which were then incubated overnight at 4°C. The plates were washed, incubated with biotin-conjugated goat anti-mouse IgG-Fc fragment antibodies (Bethyl Laboratories, Montgomery, TX, USA) for 1 h at room temperature, washed, and then incubated with horseradish peroxidase-conjugated streptavidin (BioLegend) for 30 min at room temperature. After the wash, 1×TMB substrate solution (Thermo Fisher Scientific) was added to each well, and the plates were incubated at room temperature for 15 min. Phosphoric acid (2 M) was used to stop the reaction, and the absorbance was read at 450 and 570 nm using an iMark Microplate Absorbance Reader (Bio-Rad, Hercules, CA, USA).

Urine albumin and creatinine were measured by Fuji DRI−CHEM 3500V and DRI-CHEM slides (Fujifilm).