This paper describes Sephadex G-100 chromatography of rat kidney extract containing various enzyme inhibitors. The high molecular weight renin (molecular weight above 50 000) constitutes about 50% of the total renin activity. Omission of the enzyme inhibitors yield solely low molecular weight renin. Upon rechromatography high molecular weight renin eluted in two peaks at lower molecular weight with a concomitant reduction of renin activity. Renin activity in the fractions from Sephadex G-100 chromatography was increased 70% by dialysis at acid as well as neutral pH through the whole molecular weight range. Cold storage of extract with low molecular weight increased renin activity about 25%. The results suggest that the fully active enzyme is not represented by the lower molecular weight forms of renin and direct connection between activation of renin and reduction of renin molecular size was not indicated.