Cr assay kit

Whole blood samples (20 μL) were hemodiluted with diluent NK (2 mL; Nihon Kohden Co., 8-522) and then measured using the MEK-6318K Automatic Blood Cell Analyzer (Nihon Kohden Co.). The activity/levels of serum creatine kinase (CK), creatinine (Cr), and alanine aminotransferase (ALT) were evaluated with the Beckman Au2700 Automatic Biochemical Analyzer (Beckman Coulter, Inc.) using CK assay kit (Nanjing Jiancheng Biotech. A032), Cr assay kit (Nanjing Jiancheng Biotech. C011-1), ALT assay kit (BioSino Bio-technology. 142331) The serum concentration of tumor necrosis factor alpha (TNF-α; 20140403AB5), lipopolysaccharide (LPS), interleukin 6 (IL-6; 20140421BG1), interleukin 10 (IL-10; 20140423MC1), CK-myocardial band (CK-MB; 20140421ND5), and cardiac troponin I (cTnI; 20140421MG3) was detected via the ELISA kits from Shanghai Zhuokang Biological Technology.

Blood (1.0 ml) was drawn from the jugular vein at 24 h prior to CLP (as baseline), and at 2, 8, and 24 h post-CLP. Serum was collected using EDTA as an anticoagulant. Samples were centrifuged for 15 min at 1000 × g at 2–8 °C within 30 min of collection. The supernatant was collected for assaying.
Serum Gal-3 levels were measured at baseline, 2, 8, and 24 h post-CLP, using an enzyme-linked immunosorbent assay (ELISA) (Lifespan Biosciences, Seattle, WA, USA; detection range: 0.156 ng/ml to 10 ng/ml), and serum IL-6 level was measured at baseline, 2, 8, and 24 h post-CLP, using an ELISA (R&D Systems, Minneapolis, MN, USA). Additionally, rat serum Cr levels were measured to assess renal function using a Cr assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

DFO, DFX, ferrous chloride (FeCl₂), ferric chloride (FeCl₃), and iron dextran were obtained from Sigma Aldrich (USA). Ca‐AM was purchased from the AAT bioquest (USA). CCK‐8, Cremopho EL, and MDA assay kits were purchased from Solarbio (Beijing, China). Serum iron assay kit, alanine aminotransferase assay kit (ALT), aspartate aminotransferase assay kit (AST), lactate dehydrogenase assay kit (LDH), and Cr assay kit were obtained from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). C11‐BODIPY^581/591 probes were obtained from Invitrogen (USA). FerroOrange probes were purchased from Dojindo Molecular Technologies, Inc. (Japan). Methanol and acetonitrile were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Dulbecco's modified Eagle's medium (DMEM), RPMI 1640, and phosphate buffered saline (PBS) were purchased from Corning (USA). Fetal bovine serum (FBS) was purchased from Gibco Inc. (USA).