Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker

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Publication protocol

Cultured HCECs or mouse corneal frozen sections were hydrated in PBS after fixed in 4% paraformaldehyde for 20 minutes, followed by incubation in 0.2% Triton X‐100 for 10 minutes. After rinsing three times with PBS for 5 minutes each and pre‐incubation for 1 hour with 2% bovine serum albumin at room temperature to block non‐specific staining, they were incubated with primary antibodies (anti‐LC3B and SQSTM1/p62) overnight at 4℃. After washing with PBS three times for 10 minutes each, specimens or cells were incubated with secondary antibodies for 1 hour. After washing with PBS three additional times for 15 minutes, sections or cells were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI) (Dalian Meilun Biotechnology Co., Ltd, Dalian, China) and then mounted for analysis under a Leica DM2500 microscope (Leica Microsystems, Wetzlar, Germany).

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