SQSTM1 Antibody (D-3)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
For optimal results, use low fluorescence PVDF membranes	Dilute Ab at 1:200

Upstream tips
For optimal results, use low fluorescence PVDF membranes
Protocol tips
Dilute Ab at 1:200

Publication protocol

Immunoblotting was performed as described in our previous reports.28,29 In brief, cells were lysed and the total proteins were quantified using the Bradford assay (Bio-Rad, 500-0006). Equal amounts of total protein (25 μg) in each sample were loaded onto a 15% SDS-PAGE gel. After transferring, the blot was incubated with a rabbit anti-LC3B (1:1000), a rabbit anti-MTOR (1:1000), a rabbit anti-phospho-MTOR (1:1000), a mouse anti-SQSTM1/p62 (1:200), or a mouse HRP-conjugated anti-actin (1:30000) antibody. Images were taken using a ChemiDoc™ MP System (Bio-Rad, Hercules, CA). The intensity of the band was quantified using Image Lab software (Bio-Rad) or ImageJ as described earlier. The fold change of LC3B-II is defined as the ratio of the intensity of the LC3B-II band normalized to that of actin.

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Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for SQSTM1 Antibody (D-3) below.

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