Rabbit monoclonal anti fak

Lung tissues or cell pellets were lysed in RIPA buffer containing proteinase inhibitor. Equal amounts of protein (20 μg) were loaded on 8–10% SDS-PAGE gels and then electro-transferred onto PVDF membranes (Millipore Corp). The membranes were then blocked with BSA and incubated with the indicated primary antibody (1:3,000) overnight at 4 °C. Subsequently, the membranes were incubated with a secondary antibody (1:3,000, Abcam, anti-rabbit IgG). Then, the protein level on the blot was detected using the Western Bright ECL kit (Bio-Rad Laboratories). Equal loading of the samples was validated by the detection of GAPDH. The following antibodies against the target proteins in the study were used: rabbit monoclonal anti-FAK (Abcam), rabbit polyclonal anti-phospho-FAK (at Tyr397, Cell Signalling), mouse monoclonal anti-phosphated-Akt (at Ser 473, Santa cruz), and rabbit monoclonal anti-GAPDH (Abcam).

Primary antibodies: rabbit polyclonal anti-p-FAK^Tyr397 and anti-tenascin-C (Invitrogen, Carlsbad, CA, USA); mouse monoclonal anti-osteopontin, anti-α-SMA, goat monoclonal anti-Akt1, and rabbit polyclonal anti-vitronectin (Santa Cruz Biotech., Santa Cruz, CA, USA); rabbit polyclonal anti-fibronectin (Dako, Denmark, SC, USA); rabbit monoclonal anti-FAK (Abcam, Cambridge, MA, USA); rabbit polyclonal anti-laminin, mouse monoclonal anti-α-tubulin, and anti-vinculin (Sigma-Aldrich Co., St. Louis, MO, USA); rabbit monoclonal anti-E-cadherin, anti-SMAD2, anti-p-ERK1/2^Thr202/204, anti-p-SMAD2^Ser465/467, and polyclonal anti-actin, anti-ERK 1/2, anti-SMAD4, and anti-N-cadherin (Cell Signaling Technol., Danvers, MA, USA); mouse monoclonal anti-p-Akt^Ser473, anti-actin, and rabbit monoclonal anti-histone (H3) (Millipore, Billerica, MA, USA). Secondary biotin-conjugated antibodies against rabbit IgG, mouse IgG, and goat IgG (Cell Signaling Technol. Danvers, MA, USA); Streptavidin-conjugated FITC and streptavidin-conjugated horseradish peroxidase (Carlsbad, CA, USA).