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An improved isolation procedure for carbonic anhydrase C from red cell lysate
The Tohoku Journal of Experimental Medicine
|December 1, 1980
Summary
This study details an effective method for isolating carbonic anhydrase C (CA-C) from red blood cells. The described purification process ensures high recovery and purity of the target enzyme.
Area of Science:
- Biochemistry
- Enzymology
- Protein Purification
Background:
- Carbonic anhydrase C (CA-C) is a crucial enzyme found in red blood cells.
- Efficient isolation methods are essential for studying enzyme function and developing therapeutics.
Purpose of the Study:
- To describe an effective and high-yield method for isolating carbonic anhydrase C (CA-C) from red cell lysate.
- To establish a scalable purification protocol for CA-C.
Main Methods:
- Dialysis of red cell lysate against a phosphate buffer.
- Chromatographic separation using CM-Sephadex and DEAE-cellulose columns.
- Large-scale preparation involving ethanol and chloroform treatment of the hemoglobin fraction.
Main Results:
- Complete isolation of carbonic anhydrase C (CA-C) was achieved.
- The purification method demonstrated a high recovery rate.
- Purity was confirmed using polyacrylamide gel disc electrophoresis and immunoelectrophoresis.
Conclusions:
- The described method provides an effective means for isolating carbonic anhydrase C (CA-C).
- The protocol is suitable for both small-scale purification and large-scale preparation of the enzyme.