Western Blot Analysis of Cell Lysates

For western blot analysis, total cell lysates were prepared as previously described [62 (link)]. Nitrocellulose membranes were incubated overnight with primary antibodies at 4 °C, followed by 1 h incubation with Alexa Fluor-labeled secondary antibodies (IRDye 680LT goat-anti mouse or IRDye 800CW goat anti-rabbit antibodies (LI-COR Biosciences)) at room temperature. Fluorescent images were acquired and quantified by LI-COR Odyssey Imaging System. Antibodies used are detailed in Supplementary materials and methods section.

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Reyes-Uribe P., Adrianzen-Ruesta M.P., Deng Z., Echevarria-Vargas I., Mender I., Saheb S., Liu Q., Altieri D.C., Murphy M.E., Shay J.W., Lieberman P.M, & Villanueva J. (2018). Exploiting TERT dependency as a therapeutic strategy for NRAS-mutant melanoma. Oncogene, 37(30), 4058-4072.

Publication 2018

IRDye 680LT goat-anti mouse IRDye 800CW goat anti-rabbit antibodies Odyssey Imaging System

Anti antibodies Antibodies Cell Goat Irdye 800cw Membranes Mouse Nitrocellulose Rabbit Western blot analysis

Corresponding Organization : The Wistar Institute

Other organizations : The University of Texas Southwestern Medical Center

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Variable analysis

independent variables

Primary antibodies

dependent variables

Fluorescent signal intensity

control variables

Total cell lysates preparation
Nitrocellulose membrane incubation conditions (overnight at 4 °C)
Secondary antibody incubation conditions (1 h at room temperature)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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