SQSTM1 Antibody (D-3)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Lyse cells in M-PER Mammalian Protein Extraction Reagent	- Use a dilution range 1:1001:1000

Upstream tips
- Lyse cells in M-PER Mammalian Protein Extraction Reagent
Protocol tips
- Use a dilution range 1:1001:1000

Publication protocol

Western blotting detected the protein level of MAP1LC3B, ATG12 and ATG5 in Caco-2 cells as described previously [23]. Briefly, cells were homogenized and washed with pre-cooling PBS and then lysed by the M-PER Mammalian Protein Extraction Reagent (Pierce, 78501, Thermo Scientific, Waltham, MA, USA). The protein assay kit (Pierce, 23227, Thermo Scientific) was used to measure the protein concentration. The lysates were separated by SDS-PAGE, and transferred to polyvinylidene difluoride membranes. Primary antibodies were diluted 1:1000. Membranes were developed using Supersignal® West Dura Duration substrate reagent (Thermo Scientific, 34080). Densitometric analysis on the western blot was done by Image Gauge software (Fujifilm, Maryland, USA). β-actin was used as an internal control, and the ratio of the intensity of interest protein to β-actin was calculated.

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Manufacturer protocol

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