Description:
Lung Tumor Lysate was prepared by homogenization in modified RIPA buffer (150 mM
sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM
phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1%
sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin.
Tissue and cell debris was removed by centrifugation. Protein concentration was
determined with protein assay. The product was boiled for 5 min in 1 x SDS sample
buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01%
bromophenol blue) containing 5% β-mercaptoethanol.