An assay for carbonic anhydrase activity and reactions that produce radiolabeled gases or small uncharged molecules.
An assay procedure for carbonic anhydrase activity is described. Unique to this method is the use of radiolabeled bicarbonate and a reaction vessel with a gas-permeable membrane. The method detects 14CO2 production directly and can be used at room temperature and with solutions buffered over a wide pH range. Assays are rapid (5-10 s), discounting elution and scintillation counting time. The equipment required is simple and inexpensive, marking the method ideal for routine determinations of carbonic anhydrase activity. The method can also be applied to other reactions that liberate radioactive CO2, other labeled gases, or small unchanged, labeled molecules.
Stemler
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Department of Botany, University of California, Davis 95616.
D000818:Animals; D001639:Bicarbonates; D002021:Buffers; D002245:Carbon Dioxide / Q000032:analysis*; D002250:Carbon Radioisotopes; D002256:Carbonic Anhydrases / Q000097:blood / Q000378:metabolism*; D002417:Cattle; D004912:Erythrocytes / Q000201:enzymology; D005740:Gases / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D012680:Sensitivity and Specificity; D012964:Sodium; D017693:Sodium Bicarbonate; D013997:Time Factors
D001639:Bicarbonates; D002021:Buffers; D002250:Carbon Radioisotopes; D005740:Gases; D002245:Carbon Dioxide; D017693:Sodium Bicarbonate; D012964:Sodium; D002256:Carbonic Anhydrases