Goat anti dmpk

All IHC staining was performed on formalin-fixed and paraffin-embedded tissue sections, 4μm in thickness. Sections were deparaffinized, rehydrated and endogenous peroxidase activity blocked by incubation with 3% hydrogen peroxide in 100% alcohol (1:1) for 15 min. Antigen retrieval (for CXCL1) was performed using target retrieval solution (pH 6.0; Dako) and heat induction (120°C at 15psi) for 30 sec. Sections were incubated with following primary antibodies: rabbit anti-CXCL1 (Protein Tech, 1:100), goat anti-DMPK (Santa Cruz, 1:25), and rabbit anti-Neurabin 1 (Abcam, 1:500). Exposure to primary antibodies was followed by incubation with appropriate HRP-labeled polymer secondary antibodies (Dako) and staining visualized with a 3,3′-diaminobenzidine (DAB)/substrate-chromogen system (Dako). Gill’s hematoxylin was used for counterstaining. For anti-DMPK staining sections were treated separately with avidin, biotin and 3% rabbit serum for 10 min followed by primary antibody incubation and subsequent exposure to biotinylated rabbit anti-goat secondary antibody/streptavidin-HRP for 30 min. Control tissues included macroscopically normal renal cortex taken from kidneys resected for localized neoplasia, brain, skin, intestine, heart, tonsil, skeletal muscle, and ovarian carcinoma. Semiquantitative scoring of stained kidney sections was performed in a blinded fashion.