Mouse anti human kim 1

6 µm cryosections were prepared and dried overnight prior to fixation in acetone/ethanol. Sections were blocked for 30 min in 4% milk and 15 min in 10% goat serum and incubated with primary Ab diluted in Tris-buffered saline (TBS) for 1 hour at room temperature. Primary Abs were mouse anti-human Kim-1 (Clone 219211; R&D Systems, Minneapolis, MN, USA), mouse anti-human Collagen III (Clone FH-7A; Abcam, Hong Kong) and mouse anti-human Vimentin (Clone V9; Dako, Glostrup, Denmark). Following three 5 min washes in TBS, sections were treated with 1% H₂O₂ in TBS for 10 min to block endogenous peroxidase prior to 30 min incubation in goat anti-mouse horseradish peroxidase. Sections were then washed 3 times in TBS and developed with DAB for 5 min prior to light counterstaining with haematoxylin and eosin.

Primary antibodies used in the study were: sheep‐anti mouse UMOD, goat‐anti‐mouse KIM‐1, mouse‐anti‐human KIM‐1, goat‐anti mouse NGAL (R&D Systems Inc, Minneapolis, MN, USA); sheep‐anti‐human UMOD, rat anti‐mouse F4/80 (AbD Serotec/Bio‐Rad, Mississauga, ON, Canada); mouse anti‐mouse αSMA, rabbit anti‐human fibronectin (Sigma‐Aldrich, St. Louis, MO, USA); rabbit anti‐mouse/human/rat TRPV5 (Alomone Labs, Jerusalem, Israel); mouse anti‐human PCNA (eBioscience/Thermo Fisher Scientific, Waltham, MA, USA); rabbit anti‐mouse/human/monkey megalin/LRP‐2 (Abcam, Toronto, ON, Canada); rabbit‐anti‐mouse Laminin (Millipore/Sigma), mouse‐anti‐mammalian acetylated α‐Tubulin (Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit anti‐mouse GAPDH (Cell Signaling Technology, Danvers, MA, USA).
Secondary horseradish peroxidase (HRP) conjugated anti‐IgG antibodies were: donkey anti‐sheep (Jackson ImmunoResearch Labs, West Grove, PA, USA); goat anti‐rat and horse anti‐mouse (Cell Signaling Technology); goat anti‐rabbit and donkey anti‐goat (Santa Cruz Biotechnology).
Secondary antibodies used in immunofluorescent microscopy were highly cross‐absorbed Alexa Fluor 488 or 568 conjugated antibodies (Invitrogen/ThermoFisher Scientific) for detection of mouse, rabbit, rat, sheep, and goat IgG.