Neutrophil gelatinase-associated lipocalin as a potential novel biomarker for ventilator-associated lung injury

Xiao, Rui; Chen, Rongchang

Immunohistochemical staining of NGAL. Following deparaffinization and dehydration, the sections were immersed in 10 mM sodium citrate buffer (pH 6.0) with 0.05% Tween-20, for 5 min at 58ËšC, and subsequently treated with 0.25% Triton X-100 for 5 min. Following blocking with 5% bovine serum albumin (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) at room temperature for 20 min, the slides were incubated at 4ËšC with a rat monoclonal anti-NGAL antibody (MAB1857; 1:100; R&D Systems, Inc.) overnight. Finally, slides were incubated with a rabbit anti-rat biotin-conjugated IgG H&L secondary antibody (ab6733; 1:1,000; Abcam, Cambridge, MA, USA) at room temperature for 1 h, and detected using an avidin-biotin complex system (Vectastain ABC-kit; Vector Laboratories, Inc., Burlingame, CA, USA).