The small heat shock protein 27 (Hsp27) is an oligomeric, molecular chaperone in vitro. This chaperone activity and other physiological roles attributed to Hsp27 have been reported to depend on the state of self-association. In the present work, we have used sedimentation velocity experiments to demonstrate that the self-association of Hsp27 is independent of pH and ionic strength but increases significantly as the temperature is increased from 10 to 40 ºC. The largest oligomers formed at 10 ºC are ~8-12mers while at 40 ºC, oligomers as large as 22-30-mers are observed. Similarly, the chaperone activity of Hsp27 as indicated by its ability to inhibit DTT-induced insulin aggregation also increases with increased temperature with a particularly sharp increase in activity as temperature is increased from 34 to 43 ºC. Similar studies of an Hsp27 triple variant that mimics the behavior of the phosphorylated protein establish that this protein has greatly diminished chaperone activity that responds minimally to increased temperature. We conclude that Hsp27 can exploit a large number of oligomerization states and that the range of oligomer size and the magnitude of chaperone activity increase significantly as temperature is increased over the range that is relevant to the physiological heat shock response.