The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit

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	To monitor autophagosome maturation, i.e., traffic of autophagosomes and fusion with lysosomes (also called autophagic flux), MDA-MB-231 cells were transfected as described in 6-well plates with 600 ng of a plasmid expressing full-length APC (WT or m4). 12-16 h after transfection, the medium was replaced with serum-free media (Thermo Fischer Scientific) supplemented with 20 mM HEPES (pH 7.4) and 20 mM Lglutamine. Cells were then mixed with Premo™ Autophagy Tandem Sensor RFP-GFPLC3B (#P36239; Thermo Fischer Scientific) and incubated for 16 h as described in manufacturer’s instructions.

Protocol tips

To monitor autophagosome maturation, i.e., traffic of autophagosomes and fusion with lysosomes (also called autophagic flux), MDA-MB-231 cells were transfected as described in 6-well plates with 600 ng of a plasmid expressing full-length APC (WT or m4). 12-16 h after transfection, the medium was replaced with serum-free media (Thermo Fischer Scientific) supplemented with 20 mM HEPES (pH 7.4) and 20 mM Lglutamine. Cells were then mixed with Premo™ Autophagy Tandem Sensor RFP-GFPLC3B (#P36239; Thermo Fischer Scientific) and incubated for 16 h as described in manufacturer’s instructions.

Publication protocol

To monitor autophagosome maturation, i.e., traffic of autophagosomes and fusion with lysosomes (also called autophagic flux), MDA-MB-231 cells were transfected as
described in 6-well plates with 600 ng of a plasmid expressing full-length APC (WT or m4). 12-16 h after transfection, the medium was replaced with serum-free media (Thermo Fischer Scientific) supplemented with 20 mM HEPES (pH 7.4) and 20 mM Lglutamine. Cells were then mixed with Premo™ Autophagy Tandem Sensor RFP-GFPLC3B (#P36239; Thermo Fischer Scientific) and incubated for 16 h as described in manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit below.

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