The anti-Rac1-GTP antibody and the detection of active Rac1: a tool with a fundamental flaw

Small GTPases. 2022 Jan;13(1):136-140. doi: 10.1080/21541248.2021.1920824. Epub 2021 Apr 29.

Abstract

Rac1 is a member of the Rho GTPase family and is involved in many cellular processes, particularly the formation of actin-rich membrane protrusions, such as lamellipodia and ruffles. With such a widely studied protein, it is essential that the research community has reliable tools for detecting Rac1 activation both in cellular models and tissues. Using a series of cancer cellular models, we recently demonstrated that a widely used antibody for visualizing active Rac1 (Rac1-GTP) does not recognize Rac1 but instead recognizes vimentin filaments (Baker MJ, J. Biol. Chem. 295:13698-13710, 2020). We believe that this tool has misled the field and impose on the GTPase research community the need to validate published results using this antibody as well as to continue the development of new resources to visualize endogenous active Rac1.

Keywords: Anti-Rac1-GTP antibody; Rac1; Rho GTPases; cancer cells; immunofluorescence; vimentin.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actins / metabolism
  • Guanosine Triphosphate
  • cdc42 GTP-Binding Protein* / metabolism
  • rac1 GTP-Binding Protein* / metabolism
  • rho GTP-Binding Proteins / metabolism

Substances

  • cdc42 GTP-Binding Protein
  • rac1 GTP-Binding Protein
  • rho GTP-Binding Proteins
  • Actins
  • Guanosine Triphosphate