LC3A/B (D3U4C) XP® Rabbit mAb

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Lyse cells in RIPA buffer that contained 1% protease and phosphatase inhibitors	- Dilute Ab at 1:1000 - Incubate primary antibody overnight at 4°C.

Upstream tips
- Lyse cells in RIPA buffer that contained 1% protease and phosphatase inhibitors
Protocol tips
- Dilute Ab at 1:1000 - Incubate primary antibody overnight at 4°C.

Publication protocol

Total protein was harvested using standard RIPA buffer that contained 1% protease and phosphatase inhibitors (Thermo Fisher Scientific, Canoga Park, CA, United States). Equal proteins were subjected to electrophoresis on a 8 or 12% SDS-polyacrylamide gel, then transferred into PVDF membrane. After blocked in 5% skim milk for an hour, the membrane was incubated in primary antibodies overnight at 4°C. Then, the membrane were hybridized with the secondary horseradish peroxidase-conjugated antibodies at room temperature for an hour. Finally, protein bands were visualized by the enhanced chemiluminescence (ECL) system (Millipore, United States), and the expression of protein was measured using the ImageJ software. Here, β-actin was used as an internal control.

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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for LC3A/B (D3U4C) XP® Rabbit mAb below.

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