Anti myc tag mab magnetic beads

PANC1 cells stably expressing HSP47‐Myc and SERCA2a‐Myc were lysed with a lysis buffer (50 mmol/L Tris‐HCl pH 7.5, 150 mmol/L NaCl, 1 mmol/L EDTA, 1% NP‐40 and complete mini protease inhibitors (Roche)) on ice. The soluble protein fraction was collected by centrifugation at 12 000 × g for 15 min, and protein quantification was performed using a BCA assay (Thermo Fisher Scientific). Protein aliquots each containing approximately 1 mg of cellular proteins were incubated with anti‐myc‐tag mAb‐magnetic beads (Medical & Biological Laboratories) for 16 h at 4°C, and then the beads were washed with the lysis buffer on a magnetic stand. Proteins were eluted from the beads with 50 mmol/L glycine‐HCl buffer (pH 3.5) containing SDS‐PAGE sample buffer. Eluents were immediately transferred to a fresh tube containing 5 μL of 1 M Tris‐HCl buffer (pH 7.5) in advance to neutralize sample pH.