BECN1 Antibody (H-300)

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	- Primary Ab dilution-1:1000

Protocol tips
- Primary Ab dilution-1:1000

Publication protocol

The protein extracts obtained from total cell lysate were subjected to SDS-PAGE in a Bio-Rad slab gel apparatus (Mini-PROTEAN Tetra cell; Bio-Rad, Hercules, CA, USA) and electrophoretically transferred to a nitrocellulose membrane. The membranes were incubated with primary antibodies for anti-Beclin-1 (1:1000, Santa Cruz Biotechnology, Danvers, MA, USA); anti-LC3 (1:1000, Novus Biologicals, LLC, Littleton, CO, USA); anti-p62/SQTSM1 (1:1000, Abcam, Cambridge, UK); anti-phospho-eif2α (peif2α, 1:500; Santa Cruz Biotechnology), and anti-eif2α (1:1000, Santa Cruz Biotechnology), followed by the appropriate secondary antibodies. As an internal control for protein loadings, the membranes were hybridized against β-actin (1:500, Santa Cruz Biotechnology). Immunoreactive bands were visualized using the enhanced chemiluminescence method (Super Signal CL-HRP Substrate System; Pierce Biotechnology, Rockford, IL, USA). Exposed films were scanned with a densitometer (Bio-Rad) and analyzed quantitatively with commercial equipment (Multi-Analyst Macintosh Software for Image Analysis Systems; Bio-Rad). At least three independent experiments were carried out.

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