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

Information on making liposomes from Avanti lipids web site

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