ClC-2 chloride channel is present in the brain and some transporting epithelia where its function is poorly understood. We have now demonstrated that the surface channels are rapidly internalised and approximately the 70% of the surface membrane protein recycles after 4- to 8-min internalisation. Endocytosis of ClC-2 was dependent upon tyrosine 179 located within an endocytic motif. Rapid recycling accompanied by an even faster internalisation could account for the abundant presence of ClC-2 in intracellular membranous structures. At least a proportion of ClC-2 resides in lipid rafts. Use of beta-cyclodextrin led to an increase in cell surface channel, but, surprisingly, a decrease in functionally active channels. We suggest that ClC-2 requires residing in beta-cyclodextrin sensitive clusters with other molecules in order to remain active. Regulation of ClC-2 trafficking to and within the membrane could be a means of modulating its activity. Cell. Physiol. 221: 650-657, 2009. (C) 2009 Wiley-Liss, Inc.