Anti phospho trka anti p trka

Protein extraction from cell lines and Western blotting experiments were performed as previously described [29 (link)]. Complete mini-protease inhibitor cocktail tablets (catalogue number 4693124001, Roche, Basel, Switzerland) and PhosSTOP phosphatase inhibitor tablets (catalogue number 4906837001, Roche) were also used. Samples were centrifuged at 16,000× g for 15 min at 4 °C and the supernatant was extracted and stored at −80 °C prior to Western blotting. An anti-TrkA antibody (catalogue number ANT018, Alomone Labs, Jerusalem, Israel) was used at a dilution of 1:500 and a β-actin antibody (catalogue number A2066, Sigma-Aldrich) was used at a 1:5000 dilution as the equal loading control. The following antibodies from Cell Signaling Technology were all used at a dilution of 1:500 to assess cellular protein and downstream signaling pathways: anti-HER2/ErbB2 (catalogue number 2242), anti-phospho-TrkA (anti-p-TrkA) (Tyr490, catalogue number 9141), anti-p44/42 MAPK (Erk1/2) (catalogue number 9107), anti-phospho-p44/42 MAPK (p-Erk1/2) (Thr202/Tyr204, catalogue number 4370), anti-SRC (catalogue number 2123), anti-phospho-SRC (anti-p-SRC) (Tyr416, catalogue number 2101), anti-AKT (catalogue number 2920) and anti-phospho-AKT (anti-p-AKT) (Ser473, catalogue number 4060).