LC3B antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Incubate membrane with 5% non-fat milk for 1 h at room temperature before adding primary antibody. - Dilute Ab at 1:50 and incubate for overnight at 4C.

Protocol tips
- Incubate membrane with 5% non-fat milk for 1 h at room temperature before adding primary antibody. - Dilute Ab at 1:50 and incubate for overnight at 4C.

Publication protocol

Cells were washed once with ice-cold PBS and harvested in a cell lysis buffer mixed with a protease inhibitor cocktail following various treatments. To prepare protein samples for immunoblot, the kidney tissue samples were homogenized with cell lysis buffer and with protease inhibitor cocktail. Proteins were separated by SDS/PAGE and transferred to nitrocellulose membranes. After incubation with 5% non-fat milk for 1 h at room temperature, membranes were incubated with a primary antibody overnight at 4°C and then incubated with appropriate horseradish peroxidase–conjugated secondary antibody for 1 h at room temperature. Bound antibodies were visualized by chemiluminescence detection. The densitometry analysis of immunoblot results was determined by NIH Image software (National Institutes of Health, Bethesda, MD).

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Manufacturer protocol

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