• 圣淘沙名胜世界(新加坡)

    LC3-lipidation-assay

    Liv Jensen

    Published: 2022-12-03 DOI: 10.17504/protocols.io.kqdg392ypg25/v1
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    Abstract

    Protocol for an in vitro LC3 lipidation assay using purified proteins and synthetic liposomes.

    Steps

    1.

    Mix 2μM ATG3, 2μM ATG7, 1μM WIPI2, 200nM ATG12–ATG5-ATG16L1-GFP, 5μM LC3B

    1:1 with extruded liposomes in reaction buffer.

    2.

    Incubate at 37 ̊C

    3.

    At 0, 15, 30, 60, and 120 minutes, remove 12μl of reaction mixture andquench by

    adding 4ul of SDS-PAGE loading bufferandheatingat 60 ̊C for 10 minutes.

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