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  • A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis.

A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis.

International journal of molecular sciences (2020-09-06)
Mohamad Assi, Boris Pirlot, Vincent Stroobant, Jean-Paul Thissen, Patrick Jacquemin
ABSTRACT

KRAS is a powerful oncogene responsible for the development of many cancers. Despite the great progress in understanding its function during the last decade, the study of KRAS expression, subcellular localization, and post-translational modifications remains technically challenging. Accordingly, many facets of KRAS biology are still unknown. Antibodies could be an effective and easy-to-use tool for in vitro and in vivo research on KRAS. Here, we generated a novel rabbit polyclonal antibody that allows immunolabeling of cells and tissues overexpressing KRAS. Cell transfection experiments with expression vectors for the members of the RAS family revealed a preferential specificity of this antibody for KRAS. In addition, KRAS was sensitively detected in a mouse tissue electroporated with an expression vector. Interestingly, our antibody was able to detect endogenous forms of unprenylated (immature) and prenylated (mature) KRAS in mouse organs. We found that KRAS prenylation was increased ex vivo and in vivo in a model of KRASG12D-driven tumorigenesis, which was concomitant with an induction of expression of essential KRAS prenylation enzymes. Therefore, our tool helped us to put the light on new regulations of KRAS activation during cancer initiation. The use of this tool by the RAS community could contribute to discovering novel aspects of KRAS biology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Mercury(II) amidochloride, ≥95.0%, powder
Sigma-Aldrich
FTI-277 trifluoroacetate salt, ≥95% (HPLC), film
Sigma-Aldrich
GGTI 298 trifluoroacetate salt hydrate, ≥90% (HPLC), film
Sigma-Aldrich
Monoclonal Anti-KRAS antibody produced in mouse, clone 3B10-2F2, purified immunoglobulin, buffered aqueous solution