LC3B Antibody Kit for Autophagy

Protocol tips

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	Primary antibody dilution is 1:500 for 1hour at RT

Protocol tips
Primary antibody dilution is 1:500 for 1hour at RT

Publication protocol

Autophagy was visualized using LC3B Antibody Kit for Autophagy (Invitrogen) according to the manufacturer’s instructions. Briefly, cells were grown on glass cover in 6-well plate and treated with sodium arsenite for 6 h and fixed with 3.7 % formaldehyde in PBS and permeabilized with 0.2 % Triton X-100 in PBS. Rabbit anti-LC3B primary antibody was diluted in blocking buffer (1:500) and incubated 1 h at room temperature. After washing three times with PBS, cells were incubated with anti-rabbit secondary antibody followed by mounting with PI. Image was captured using a Nikon confocal microscope (Nikon TE200-C1) at 24 °C room temperature. Number of autophagosome positive cell was counted. In addition, LC3B protein expression was measured with Western blotting.

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