Anti-LC3 antibody produced in rabbit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Lyse cells in Laemmli SDS buffer	Incubate primary Ab with agitation for 1 hour. The solution should move freely across the surface of the membrane.

Upstream tips
Lyse cells in Laemmli SDS buffer
Protocol tips
Incubate primary Ab with agitation for 1 hour. The solution should move freely across the surface of the membrane.

Publication protocol

After the designated treatments, Laemmli SDS buffer were used to lyse the collected samples (62.5 mM Tris, pH 6.8, 25% glycerol, 2% SDS [Vivantis, PB0640], phosphatase inhibitor [Thermo Scientific, 78428] and proteinase inhibitor cocktail [Roche, 11697498001]). An equal amount of protein of each sample was resolved by SDS-PAGE, and then the SDS-PAGE gels were transferred onto PVDF membrane (Bio-Rad, 162–0177). Membranes were blocked for 30 min with blocking buffer (Thermo, 37538) and followed by incubations with the indicated primary and secondary antibodies. Then the membranes were visualized with the enhanced chemiluminescence method (Thermo Scientific, 34076) by the ImageQuant LAS 500 (GE).

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Manufacturer protocol

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