Rabbit anti mouse itln antibody

Mouse lungs and BEAS-2B cells were harvested at the indicated time points, twice with cold PBS, and lysed in RIPA lysis buffer (Goodbio technology, China) containing a protease inhibitor cocktail (Roche, Germany). After 30 min on ice, lysates were centrifuged at 15,000×g for 5 min to remove insoluble material. 50 μg protein samples were resolved on 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred and immunoprobed with rabbit-anti-mouse Itln antibody (1:1000;Proteintech Group). Antibody was detected using horseradish peroxidase-conjugated anti-rabbit IgG (1:3000; Jackson ImmunoResearch Laboratories) followed by ECL Western blot detection reagent (Beyotime Biotech, China). Blots were stripped and then re-probed for β-actin (1:3000; Abcam, Cambridge, UK). Densitometry was performed using ImageJ (National Institutes of Health, USA) and the protein level of intelectin was indexed to beta-actin. For phosphorylated EGFR, total EGFR, phosphorylated ERK and total ERK Western blotting, rabbit monoclonal phospho-EGFR (Tyr1068) (1:1000), EGFR (1:1000), phospho-p44/42 ERK1/2 (Thr202/Tyr204) (1:2000) and ERK1/2 antibody (1:1000) from Cell Signaling Technology were used.