The Wilbur-Anderson Carbonic Anhydrase activity assay is limited to an operating temperature of 4�C in order to keep the enzymatic reaction so slow that it can be measured visually by a colorimetric change or by titration in a pH meter.At higher temperatures analysis can be done by exploiting the esterase activity and using e.g. p-nitrophenyl substrates like p-nitrophenyl acetate. This requires very pure Carbonic Anhydrase preparations in order to avoid interference from esterase's present (e.g. proteases and lipases). For routine Carbonic Anhydrase analysisby Wilbur-Anderson the temperature limit can be overcome by using a 96-well plate reader like the Biotek Synergy 2 reader equipped with a dispenser unit. The dispenser unit ensures a very fast mixing with enzyme and substrate and makes it possible to use analysis temperatures up to 30�C.

Determination of carbonic anhydrase activity by Wilbur-Anderson in a 96-well plate reader

243rd National Meeting (2012)

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