Anti kras ab55391

In total, 5 × 10⁵ cells of each cell line were seeded and cultivated overnight, treated with cetuximab-protamine (60 nM) coupled to the indicated siRNAs at 1:10 molar ratio once a day for 3 days, harvested, lysed in RIPA buffer and cleared by ultrasonification and centrifugation. Xenograft tumours were homogenised as 10% w/v in RIPA buffer using an ultraturrax, cleared by ultrasonification and centrifugation. Western blot analysis was performed using standard protocols with the following antibodies: anti-KRAS (ab55391, Abcam, Cambridge, UK), and anti β-Actin antibody (Clone AC-15, Sigma Aldrich).

5x10⁵ cells of each cell line were seeded and cultivated overnight, treated with cetuximab-protamine (60 nM) coupled to the indicated esiRNAs at 1:10 molar ratio once a day for 72 h, harvested, lysed in RIPA buffer and cleared by centrifugation. Xenograft tumors were homogenized as 10% w/v in RIPA buffer using an ultraturrax, cleared by ultrasonification and centrifugation. Western blot analysis was performed using standard protocols with the following antibodies: anti-KRAS (ab55391, ABCAM, Cambridge, UK), PI3Kinase- p110α (4249), anti-ERK1/2 (4696), anti-phospho-ERK1/2 (4370), p-AKT-S473 (4058), anti-AKT, anti c-MYC (9402, all Cell Signaling Technology, Danvers, MA, USA), and anti β-Actin antibody (Clone AC-15, Sigma Aldrich). Densitometric analysis of gel-electrophoretic bands was carried out using the NIH Image J package (http://rsb.info.nih.gov/ij/).