Anti-LC3B antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Use a working concentration of 1-2 mg/mL. - Incubate primary antibody overnight at 4C.

Protocol tips
- Use a working concentration of 1-2 mg/mL. - Incubate primary antibody overnight at 4C.

Publication protocol

After respective treatment, cells were harvested and lysed in the radio immunoprecipitation assay (RIPA) buffer supplemented with protease inhibitor cocktail. Cell lysates were centrifuged, and protein concentration was determined by BCA protein assay kit (Thermo Fisher Scientific-Pierce). Equal amounts (20 µg) of total protein were subjected to SDS-PAGE and electrotransferred to PVDF membranes. Membranes were incubated overnight at 4 °C with the primary antibodies and 1 hr with the corresponding secondary antibodies, and antibody reactions were visualized with an enhanced chemiluminescence kit (Thermo Fisher Scientific-Pierce). The signals were detected and quantified using Kodak 4000 image station and companion software. The density of each target band was normalized to that of the loading control (β-actin). Data obtained were expressed as the ratio of the intensity of the protein in the treated cells to that of the corresponding protein in control cells. Each test was performed in four different experiments with different batches of cells.

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Manufacturer protocol

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