Anti-LC3B antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Incubate and dilute Primary Ab in 5% BSA in PBST overnight at 4°C

Protocol tips
- Incubate and dilute Primary Ab in 5% BSA in PBST overnight at 4°C

Publication protocol

Cells treated with reagents for indicated times were lysed in RIPA buffer supplemented with a proteinase inhibitor (10 mg/mL aprotinin, 10 mg/mL phenyl-methylsulfonyl chloride, and 50 mM sodium orthovanadate). Cell extracts were quantified using the BCA protein assay kit, and equal amounts of proteins were separated by SDS-PAGE. Gels were then transferred to PVDF membranes, blocked with 5% non-fat dry milk, and incubated with the primary antibody solution. Alternatively, primary antibodies were diluted in 5% BSA in PBST overnight at 4°C. The membrane was washed with PBST and incubated with secondary antibody conjugated with infrared dyes for 2 h at room temperature. Finally, the protein bands were detected by Odyssey infrared image system (LI-COR, Lincoln, NE, USA), and the protein expression levels were quantified by Image Studio software (Lincoln, NE, USA).

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