Part I: Generic Ras Biology
1. Ras Variant Biology and Contributions to Human Disease
Ian A. Prior
2. Regulation of the Small GTPase Ras and its Relevance to Human Disease
Kayla R. Kulhanek, Jeroen P. Roose, and Ignacio Rubio
Part II: Methods in Ras Biochemistry
3. Precise Characterization of KRAS4B Proteoforms by Combining Immunoprecipitation with Top-Down Mass Spectrometry
Lauren M. Adams, Caroline J. DeHart, and Neil L. Kelleher
4. Absolute Quantitation of GTPase Protein Abundance
Fiona E. Hood, Yasmina M. Sahraoui, Rosalind E. Jenkins, and Ian A. Prior
5. Validation of Isoform- and Mutation-specific RAS Antibodies
Andrew M. Waters and Channing J. Der
6. Production and Membrane Binding of N-terminally Acetylated, C-terminally Farnesylated and Carboxymethylated KRAS4b
Simon Messing, Constance Agamasu, Matt Drew, Caroline DeHart, Andrew G. Stephen, and William K. Gillette
7. Active GTPase Pulldown Protocol
Martin J. Baker and Ignacio Rubio
8. Methods to Monitor Ras Activation State
Kari Kopra and Harri Härmä
9. NMR Detection Methods for Profiling RAS Nucleotide Cycling
Ryan C. Killoran and Matthew J. Smith
Part III: Methods in Ras Processing, Trafficking, and Localization
10. Ras Diffusion and Interactions with the Plasma Membrane Measured by FRAP Variations Orit Gutman, Marcelo Ehrlich, and Yoav I. Henis
11. Understanding Ras Spatial Cycles through Reaction-Diffusion Simulations
Malte Schmick and Philippe I. H. Bastiaens
12. Super-Resolution Imaging and Spatial Analysis of RAS on Intact Plasma Membrane Sheets
Yong Zhou and John F. Hancock
13. FLIM-FRET Analysis of Ras Nanoclustering and Membrane-Anchorage
Hanna Parkkola, Farid Ahmad Siddiqui, Christina Oetken-Lindholm, and Daniel Abankwa
14. Assessment of Plasma Membrane Fatty Acid Composition and Fluidity Using Imaging Flow Cytometry
Natividad R. Fuentes, Michael L. Salinas, Xiaoli Wang, Yang-Yi Fan, and Robert S. Chapkin
15. Spatiotemporal Imaging of Small GTPase Activity using Conformational Sensors for GTPase Activity (COSGA)
Yao-Wen Wu
Part IV: Methods in Ras Signaling and Inhibition
16. Using BioID to Characterize the RAS Interactome
Hema Adhikari and Christopher M. Counter
17. Probing RAS function with Monobodies
Imran Khan and John P. O'Bryan
18. Detection of Endogenous RASSF1A Interacting Proteins
Howard Donninger, Desmond Harrell-Stewart, and Geoffrey J. Clark
19. Mathematical Modeling to Study KRAS Mutant-Specific Responses to Pathway Inhibition
Edward C. Stites
20. A Facile Method to Engineer Mutant Kras Alleles in an Isogenic Cell Background
Konstantin Budagyan and Jonathan Chernoff
21. RASless MEFs as a Tool to Study RAS-dependent and -independent Functions Carmen G. Lechuga, Marina Salmón, Guillem Paniagua, Carmen Guerra, Mariano Barbacid, Matthias Drosten
Part V: Methods in vivo Ras Biology
22. Generation of Patient-derived Colorectal Cancer Organoids for RAS Studies
Dirk Schumacher, Joseph L. Regan, Dorothea Przybilla, and Reinhold Schäfer
23. Ras-GEF Mouse Models for the Analysis of Ras Biology and Signaling
