Supplementary Figure 1: Western blot validation of a RAS-GTP ELISA. | Nature Cell Biology

Supplementary Figure 1: Western blot validation of a RAS-GTP ELISA.

From: RAS nucleotide cycling underlies the SHP2 phosphatase dependence of mutant BRAF-, NF1- and RAS-driven cancers

Supplementary Figure 1

NCI-H358 cells were grown in 2D culture and incubated with increasing concentrations of RMC-4450 for one hour. a) Cellular lysates were prepared and levels of RAS-GTP were measured by ELISA. Shown are three independent biological replicates performed in technical duplicate. b) Assessment of RAS-GTP in samples from replicate 2 by western blot. RAS-GTP was affinity purified from the cellular lysates using the GST-RAF-RBD bound to glutathione resin. Source data is provided in Supplementary Table 9.

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