Lab protocols:

Dynamin GTPase Assay

Rat Brain CCV prep

Protonation of phosphoinositol lipids

Making liposomes and lipid tubules

Lipid Monolayer Assay

Negative staining protocol for EM grids

We apply a variety of methods to the study of endocytosis. Some of our favorites include:

X-ray crystallography In collaboration with the laboratory of Phil Evans, the group has solved the structures of a number of endocytic proteins and domains.  See the structures of some endocytic proteins

Isothermal titration calorimetry to quantitate protein-protein interactions

Lipid Monolayer assay to reconstitute early steps of endocytosis

Overexpression of endocytic proteins in COS cells to test the effects of mutations in vivo

Liposome/lipid nanotube sedimentation assays to test lipid binding and specificity

(also immunoprecipitations, cloning, kinase assays, etc.)

Eva Schmid's methods from thesis: HeLa cell extracts, Dictyostelium methods, Yeast-2-hybrid and Surface Plasmon Resonance.

Transferrin endocytosis assay for cultured cells