Sevrin_CellRep_2024

Published: 13 August 2024| Version 1 | DOI: 10.17632/6nmytyss4n.1
Contributor:
Thomas Sevrin

Description

Original Western Blot images from all figures of Sevrin et al. 2024 Cell Report paper.

Files

Steps to reproduce

Protein concentration of cell lysates was determined using the BCA assay according to the manufacturer’s instructions (PierceTM BCA Protein Assay Kit). Loading solutions were prepared at 1 µg protein/µL using 4X Laemmeli buffer and DTT. Ten micrograms of protein were resolved by SDS PAGE using 10% acrylamide gels in Mini-PROTEAN Tetra cell (Biorad) systems. Proteins were then transferred into PVDF membranes using the XCell IITM wet transfer system (Invitrogen). Membranes were blocked for 1h at room temperature using 4% BSA in TBS-Tween 0.1% (TBS-T), then incubated with primary antibody (diluted 1:1000 in 1% BSA in TBS-T) from 1h at room temperature to overnight at 4°C. Membranes were then incubated for 1h at room temperature with HRP-coupled secondary antibodies diluted at 1:5000 in 1%BSA in TBS-T. Finally, blots were developed using homemade ECL reagent or SuperSignal™ West Femto reagent (Thermo Scientific) with the iBright™ CL750 Imaging System (Thermo Scientific).

Institutions

University College Dublin, Systems Biology Ireland

Categories

Drug Resistance in Cancer, Pancreatic Cancer, Western Blot, Targeted Therapy

Funding

National Cancer Institute

R01CA244660

Horizons Foundation

101136926

Science Foundation Ireland

22/PATH-S/10875

Licence