Anti-LC3B antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Incubate Primary antibody at 4C overnight. - If there is high background or non specific bands dilute the secondary antibody in buffer containing 1-5% normal serum from the same species as the sample

Protocol tips
- Incubate Primary antibody at 4C overnight. - If there is high background or non specific bands dilute the secondary antibody in buffer containing 1-5% normal serum from the same species as the sample

Publication protocol

After cell fraction preparation and protein quantification with a bicinchoninic acid (BCA) protein assay kit (Beyotime, Shanghai, China), equal amounts of protein were separated by 8–15% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to 0.22-μm or 0.45-μm polyvinylidene difluoride (PVDF) membranes. After transfer, membranes were blocked in 5% skim milk for 1 h at room temperature. The membranes were incubated overnight at 4 °C with the relevant primary antibodies. Membranes were then incubated with the appropriate secondary antibodies (1:5000) for 1 h at room temperature. Western blots were developed using enhanced chemiluminescence reagent. Protein levels were determined by computer-assisted densitometric analysis (GS-800 densitometer, Quantity One; Bio-Rad). The band volumes were determined by standard scanning densitometry with normalization of densitometry measures to β-actin. Each test was performed in triplicate.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Anti-LC3B antibody produced in rabbit below.

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