Anti icam

Lung homogenates were subjected to 4–15% gradient gel. Gels were transferred to PVDF membranes (Millipore) and the membranes were blocked with 3% nonfat dry milk in TBS-T (100 mM Tris (pH 7.5), 150 mM NaCl, 0.1% Tween 20) for 1 h. Proteins were detected using goat polyclonal anti-VCAM (R&D Systems, Minneapolis, MN) and anti-ICAM (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), and anti-actin-HRP (Abcam, Cambridge, MA).

Anti-GAPDH and anti-rabbit were purchased from Sigma-Aldrich Co. (St. Louis, MO, United States). DMEM/F12 culture medium, fetal bovine serum (FBS), and trypsin-EDTA were purchased from Gibco BRL (Carlsbad, United States). Anti-ICAM and anti-VCAM were purchased from Santa Cruz Biotechnology (Dallas, United States). Anti-RP-1 was obtained from BD Biosciences (San Jose, CA, United States). Histopaque-1083 and histopaque-1119 were purchased from Sigma-Aldrich Co. (St. Louis, MO, United States). Organic and inorganic compounds, salts, acids, and solvents were purchased from Merck (Darmstadt, Germany). LPS (Ultra pure lipopolysscharide from E Coli 0111:B4 strain-TLR4 ligand) Cat. code TLRL pelps, was purchased from Invivogen (San Diego, Ca, United States), TAK-242, and Collagenase type II were purchased from Invivogen (San Diego, United States). Sterile plastic material for obtaining and culturing cells was purchased from Falcon (Ballerica, Ca, United States). Xylazine 2% was purchased from Laboratorios Centrovet (Santiago, Chile). Ketamine 100 mg/ml was purchased from Richmond Vet Pharma (Bs. As., Argentina). Interferon-beta (IFN-β) was purchased from InterferonSourse (NJ, United States). MACS^® culture medium for neutrophil isolation was purchased to Miltenyi Biotec (Teterow, Germany).