Cells were lysed in RIPA lysis buffer (50 mM Tris-HCl pH 8, 150 mM NaCl, 1% v/v Nonidet P-40, 0.1% w/v SDS, 0.5% w/v sodium deoxycholate, 5 mM NaF, 5 mM β-glycerophosphate, 30 μg/ml RNase, 30 μg/ml DNase I, 1x Protease Inhibitor Cocktail (Melford Laboratories), 1 mM PMSF) followed by analysis by SDS-PAGE and Western blotting. Primary antibodies used were against: HA (1:1000, Sigma H3663), α-tubulin (1:5000, mouse clone B512, Sigma T5168), α-tubulin (1:5000, rabbit antibody, Abcam ab15246), ALK (1:1000, Cell Signalling Technology 3633), ERK (1:2000, Cell Signalling Technology 9102), and pERK (1:1000, Cell Signalling Technology 9101). Secondary antibodies used were horseradish peroxidase (HRP)-labelled IgGs against rabbit (1:2000; Sigma A6154) and mouse (1:2000, Bethyl Laboratories A90-116P) antibodies. Protein bands were visualised by enhanced chemiluminescence (ECL) according to manufacturer’s instructions (Pierce).
Ab15246
Manufactured by Cell Signaling Technology
Ab15246 is a primary antibody product manufactured by Cell Signaling Technology. It is a tool for the detection and analysis of target proteins in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ab15246
1
Protein Extraction and Western Blot Analysis
2
Cell Lysis and Protein Detection
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Cells were lysed in RIPA lysis buffer [50 mmol/L Tris-HCl pH 8, 150 mmol/L NaCl, 1% v/v Nonidet P-40, 0.1% w/v SDS, 0.5% w/v sodium deoxycholate, 5 mmol/L NaF, 5 mmol/L β-glycerophosphate, 30 µg/mL RNase, 30 µg/mL DNase I, 1× Protease Inhibitor Cocktail (Melford Laboratories), 1 mmol/L PMSF] followed by analysis by SDS-PAGE and Western blotting. Primary antibodies used were against: HA (1:1,000, Sigma H3663), α-tubulin (1:5,000, mouse clone B512, Sigma T5168), α-tubulin (1:5,000, rabbit antibody, Abcam ab15246), ALK (1:1,000, Cell Signaling Technology 3633), ERK (1:2,000, Cell Signaling Technology 9102), and pERK (1:1,000, Cell Signaling Technology 9101). Secondary antibodies used were horseradish peroxidase (HRP)-labeled IgGs against rabbit (1:2,000; Sigma A6154) and mouse (1:2,000, Bethyl Laboratories A90–116P) antibodies. Protein bands were visualized by enhanced chemiluminescence (ECL) according to manufacturer's instructions (Pierce).
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