Abstract
An effective isolation of carbonic anhydrase C (CA-C) from red cell lysate was described. The lysate dialyzed against 0.0175M phosphate buffer pH 6.3 was applied onto a CM-Sephadex column equilibrated with the same buffer. The elution was performed with the starting buffer and 0.1M dibasic potassium phosphate containing 0.14M NaCI. The hemoglobin fraction eluted with the second eluant was applied onto a DEAE-cellulose column and eluted with 0.2M glycine containing 0.01 percent KCN, resulting in complete isolation of CA-C at high recovery rate. For the preparation in a large scale, the hemoglobin fraction prepared from CM-Sephadex semibatch-type chromatography was treated with cold ethanol and chloroform. The purity of these preparations was confirmed by polyacrylamide gel disc electrophoresis and immunoelectrophoresis.
