Tim 1 kim 1

Kidneys were perfused via the left ventricle with 0.9% saline solution at 37.0 °C until the kidney cortex completely cleared of blood. Kidneys were then removed immediately after the perfusion ended and either fixed and embedded in paraffin; or shock frozen. Paraffin sections were stained with H&E and PAS for histological examination. To demonstrate the tubular damage in response to the bilateral renal ischemia, immunohistochemical staining for KIM-1 was performed on alternate sections using goat polyclonal antibody (TIM-1/KIM-1, catalogue number: AF1817, R&D Systems, Minneapolis, MN, USA) and a Vectastain Elite ABC kit (Vecor, Burlingame, CA, USA) was for visualization.

Recombinant human PSA, CA125 and KIM-1 were purchased from R&D Systems as carrier free human kallikrein 3/PSA, CA125/MUC-16, or TIM-1/KIM-1/HAVCR proteins. An ELISA for PSA was constructed using components from human kallikrein 3/PSA DuoSets (DY1344), CA125/MUC-16 DuoSets (DY 5606 or TIM-1/KIM-1/HAVCR DuoSets (DY 1715B) from R&D Systems per directions for ELISA for a fluorescent linked immunosorbent assay (FLISA) (IRDye 800CW from LI-COR Bioscience) and reformatting these directions using plasmonic-fluor as the reporter for p-FLISA (plasmonic-fluor–800CW). Streptavidin-HRP and TMB substrate were purchased from R&D System. zinc nitrate, 2-imidazolecarboxaldehyde (ICA), 2-methyimidazol (2-MI), ethylenediaminetetraacetic acid (EDTA), Polyvinylpyrrolidone (PVP), L-Cysteine, Tween-20, sodium phosphate monobasic, sodium phosphate dibasic, and 10X phosphate buffered saline (PBS) powder to make 1X PBS were purchased from Sigma-Aldrich.