Anti-LC3 pAb

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	1:2000 dilution of Primary antibody

Protocol tips
1:2000 dilution of Primary antibody

Publication protocol

Cell lysates were prepared using a radio immunoprecipitation assay (RIPA) buffer (25 mM Tris–HCl, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 0.1% sodium dodecyl sulfate; pH 7.6) containing protease inhibitors (Cocktail Tablet; Roche Diagnostics). The protein concentration of each sample was determined using a Bio-Rad protein assay kit (Bio-Rad). Identical amounts of proteins were electrophoresed in 10% or Any kD Mini-PROTEAN TGX Precast Gel (Bio-Rad) and transferred to a nitrocellulose membrane. Blots were incubated with either a rabbit EFTUD1 antibody (1:500), a mouse anti-β-actin antibody (1:4,000; Sigma), a rabbit anti-p62 antibody (1:5,000; Sigma), a horseradish peroxidase (HRP)-conjugated anti-p53 (DO-1) antibody (1:5,000; Santa Cruz Biotechnology), a rabbit anti-p21(C-19) antibody (1:1,000; Santa Cruz Biotechnology), or a rabbit anti-LC3 antibody (1:2,000; PM036; MBL). The blots were incubated with HRP-conjugated secondary antibodies (1:20 000; anti-rabbit; MBL International; anti-mouse; GE Health Care Biosciences), and were then exposed to x-ray films.

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