Anti vcam 1 antibody sc 8304

Lungs were fixed in 4% PFA for 2 hours and placed in a 30% PBS sucrose solution overnight at 4 °C before being embedded in a cutting medium and frozen at −80 °C. Serial 8-µm transversal cryo-sections of lungs were obtained. VCAM-1 expression was analyzed using rabbit polyclonal anti-VCAM-1 antibody (sc-8304) from Santa Cruz Biotechnology. For detection, horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody was used (Thermo Fisher Scientific). Sections stained with secondary antibodies alone were used as controls of non-specific staining. The slides were examined under a light microscope. Lung sections were further counterstained with hematoxylin and eosin and examined at 20× or 63× magnification using a DMi8 advanced fluorescence microscope (Leica Microsystems, Germany). NIH ImageJ software version 1.45 was used for quantification.

The chemicals used in the present study were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against ICAM-1 (Cat. no. 4915), p65 (Cat. no. 8242), lamin A/C (Cat. no. 2032) and β-actin (Cat. no. 4967) were obtained from Cell Signaling Technology (Beverly, MA, USA). Anti-VCAM-1 antibody (sc-8304) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).