SQSTM1 monoclonal antibody (M01), clone 2C11

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- primary antibody dilution immunostaining (1:100 dilution) and immunoblotting (1:2000 dilution). - incubate with the primary antibody at 4 °C overnight

Protocol tips
- primary antibody dilution immunostaining (1:100 dilution) and immunoblotting (1:2000 dilution). - incubate with the primary antibody at 4 °C overnight

Publication protocol

AML12 cells were seeded on sterile glass coverslips placed in 24-well plates at a density of 1.0 × 105 cells per well. After desired treatment, cells on coverslips were washed with PBS and fixed in 4% PFA for 8 min at room temperature. Fixed cells were washed with PBS and incubated with the following solution (1% Trion × 100, 15% goat serum, 15% 1 M Glycine diluted in water, 69% PBS) for 1 h at room temperature. The slides were then incubated with the primary antibodies at 4 °C overnight. After washing, the slides were incubated with the respective secondary antibodies diluted in PBS buffer for 1 h at room temperature. In the case of dual immunofluorescence staining, each set of primary and secondary antibodies was applied sequentially. When co-staining for the lipid droplets, the lipophilic dye, Bodipy-581/591 (1 µM in PBS) was applied at the end of the procedure for 10 min at room temperature. The slides were then mounted with ProLong® Gold Antifade Mountant, examined under the Olympus FV1000-MPE laser scanning confocal microscope. At least 50 cells from replicated cultures were randomly selected, and percentages of LDs with colocalized signals were quantified.

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Manufacturer protocol

Download the product protocol from Abnova for SQSTM1 monoclonal antibody (M01), clone 2C11 below.

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