Labeling Acetylcholine Receptors in Live Cells Using Rhodamine α-Bungarotoxin for Imaging
This protocol was adapted from “Nonimmunological Fluorescent Labeling of Cellular Structures,” Chapter 5, in Basic Methods in Microscopy (eds. Spector and Goldman). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2006.INTRODUCTION
The eukaryotic cell has evolved to compartmentalize its functions and transport various metabolites among cellular compartments. Therefore, in cell biology, the study of organization and structure/function relationships is of great importance. Integral membrane protein receptors can be studied using specifically fluoresceinated but still functional hormones or toxins that bind to specific receptors. α-bungarotoxin binds with high affinity to the α-subunit of the nicotinic acetylcholine receptor of the neuromuscular junction. In this article, acetylcholine receptors are labeled with α-bungarotoxin conjugated to rhodamine.
