MAP LC3β Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
MAP LC3β Antibody (G-9) is a mouse monoclonal IgG2b (kappa light chain)	1:300 dilution of primary antibody and incubation for 1 hour at RT

Upstream tips
MAP LC3β Antibody (G-9) is a mouse monoclonal IgG2b (kappa light chain)
Protocol tips
1:300 dilution of primary antibody and incubation for 1 hour at RT

Publication protocol

Cells were seeded into six-well plates at a density 1.0–2.0×106 cells/well and treated with alisertib (0, 0.1, 1 and 5 µM) for 24 h. Cells were then fixed with 4% paraformaldehyde (Solarbio Science and Technology, Ltd., Beijing, China) for 30 min at room temperature and permeabilized using pre-cooled methanol on ice for 10 min. Binding sites were then blocked with 5% normal goat serum (Thermo Fisher Scientific, Inc.) and 0.3% Triton X-100 (Thermo Fisher Scientific, Inc.) for 1 h. Cells were then incubated with mouse monoclonal anti-light chain (LC)3B (cat. no. sc-376404; 1:300; Santa Cruz Biotechnology, Inc.) followed by 1 h of incubation with Alexa Fluor 488 goat anti-mouse immunoglobulin G (cat. no. 8878; 1:1,000 dilution; Cell Signaling Technology, Inc., Danvers, MA, USA) as a secondary antibody. After incubation, cells were washed with ice-cold PBS, mounted onto microscopic slides with Flouromount-G (Southern Biotech, Birmingham, AL, USA) and observed using a confocal laser scanning microscope (TCS SP5; Leica Microsystems GmbH, Wetzlar, Germany).

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Manufacturer protocol

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