Anti-LC3 pAb

Protocol tips

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	For Co-IP, Use non-denaturating lysis solution to lyse the cells and incubate the cell lysate for 3 hrs at 4°C with Anti LC3 Antibody.Protein A/G then can be incubated overnight.

Protocol tips
For Co-IP, Use non-denaturating lysis solution to lyse the cells and incubate the cell lysate for 3 hrs at 4°C with Anti LC3 Antibody.Protein A/G then can be incubated overnight.

Publication protocol

The cells were then treated with SFN-NAC (30 μM) for 24 h after being seeded into 10-cm cell culture dishes and being cultured for 24 h at a density of 5 × 106 cells/dish. Next, the cells were washed three times with ice-cold PBS and lysed on ice using a non-denaturing lysis solution containing a protease inhibitor cocktail. Monoclonal anti-alpha-tubulin (Santa Cruz Biotech) or monoclonal anti-LC3 (Medical & Biological Laboratories) was added to the protein lysate and incubated for approximately 3 h at 4°C. The protein A/G agarose was then co-incubated overnight, and the purified protein complexes were recovered via centrifugation and heated to 95°C for 5 min. The protein complex indicated the binding protein (using western blot analysis).

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