Preparation and Loading of Protein Samples for Microinjection
This protocol was adapted from “Microinjection of Fluorophore-Labeled Proteins,” Chapter 5, in Live Cell Imaging (eds. Goldman and Spector). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.INTRODUCTION
This protocol provides methods for the preparation of protein samples and for loading them into pulled microinjection pipettes. Stock solutions of proteins are thawed, diluted (if desired), centrifuged at high speed to remove aggregates, and kept on ice until loading. Loading into micropipettes can be done using either a “front-loading” or a “backfilling” procedure; both methods are described below.
