Tibc assay kit

The concentrations of hemoglobin in the serum were determined using an automated hematology analyzer (Sysmex K-1000D, Sysmex Inc., Kobe, Japan). The concentrations of serum iron and total iron binding capacity (TIBC) in the serum were measured using commercial kits (serum iron assay kit, No: A039-1; TIBC assay kit, No: A040, Jiancheng, China). The concentrations of hepcidin in the serum and liver were measured by using a hepcidin ELISA kit (NO: H252, Jiancheng, China). Iron concentrations in the tissues and digesta were measured by inductively coupled plasma atomic emission spectroscopy (ICP-AES) according to the procedures described previously [17 (link)]. Briefly, 0.25 g of tissues or digesta (dried samples, the tissues and digesta were put in the freeze dryer for 24 h), 5 mL of 65% HNO₃ and 2 mL of 30% H₂O₂ were mixed in a polytetrafluoroethylene (PTFE) tube. Then, the samples were digested in a microwave sample digestion system (MARS 6, CEM Corporation, Matthews, NC, USA). Afterwards, the PTFE tubes were kept in a heater to remove acid. Five percent of HNO₃ was used to dissolve the tube residues in the PTFE tubes. Finally, the prepared solution was stored in a refrigerator at 4 °C until analysis.

The hemoglobin (Hb), serum iron (SI) concentration, and total iron binding capacity (TIBC) levels were measured using an Hb test solution, SI assay kit, and TIBC assay kit, respectively (Nanjing Jiancheng Bioengineering Inst., Nanjing, China).