SQSTM1/p62 (D1Q5S) Rabbit mAb

Protocol tips

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

Upstream tips
- Lyse cells with RIPA buffer that contained 1% protease and phosphatase inhibitors.
Protocol tips
- 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 SQSTM1/p62 (D1Q5S) Rabbit mAb below.

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