Anti-LC3B antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Incubate primary antibody overnight at 4 °C	- If no signal or weak signal or non-specific bands, make sure Enzyme inhibitor (Sodium azide) is not present as it inhibits

Protocol tips
- Incubate primary antibody overnight at 4 °C
Downstream tips
- If no signal or weak signal or non-specific bands, make sure Enzyme inhibitor (Sodium azide) is not present as it inhibits

Publication protocol

Whole cell lysates were prepared using cell lysis buffer (CST, 9803) supplemented with protease inhibitors while the subcellular fractionation was performed using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, 78,833) according to the instructions. Proteins were separated on 8–13% SDS-PAGE gel and transferred to PVDF membranes. Then the membranes were incubated with appropriate primary antibodies overnight at 4 °C and horseradish peroxidase-conjugated suitable secondary antibodies for 1 h at room temperature. The protein bands were detected with enhanced chemiluminescence (Merck Millipore, Billerica, MA, USA, WBKLS0500).

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Manufacturer protocol

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