Product: True-Phos™ Perm Buffer

Catalog No.: 425401

Gary Warnes, Dr Blizard Institute, Queen Mary University London

Use of True-Phos™ Perm Buffer Cat#76344 LO#B223447 in the Detection of Autophagy & ER Stress
Overall:	Product Quality:	Ease of Use:
The BioLegend Fix & Tru-Phos™ reagents detect autophagy and ER stress markers.

Experimental Design

Application:	Flow Cytometric Detection of LC3B and PERK
Cells used:	Jurkat Cells
Brief Protocol:	Cells after Thapsigargin treatment and labelled with Zombie NIR for 15 min at RT. Cells were washed in PBS/0.1% BSA and then fixed in 1% BioLegend Fixative for 20 mins at RT. After washing in PBS cells were treated with 100ul of True-Phos Perm for 15 min at RT. After washing cells were incubated with anti-LC3B and anti-PERK for 20 min at RT. After washing cells were labelled with Alexa Fluor® 647 * Pe-Cyanine7 for 20 min at RT. After washing cells were labelled with DAPI for cell cycle analysis. 50,000 events were collected flow cytometrically.
Results Summary:	Up-regulation of LC3B and PERK above controls were detected in a cell cycle dependent manner.
Additional Notes:

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Publication Status This data has not been published.

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