, Volume 11, Issue 2, pp 112–128 | Cite as

Histochemical demonstration of carbonic anhydrase activity

  • Holger P. J. Hansson


Freeze-dried frozen sections are floated on the surface of the freshly prepared incubation mixture (CoSO4 1.75 × 10−3M, H2SO4 5.3 × 10−2M, NaHCO3 1.57 × 10−2M and KH2PO4 1.17 to 11.7 × 10−3M; demonstration of weak activity requires high phosphate). A compound containing cobalt and phosphorous precipitates at carbonic anhydrase sites and is converted to CoS. Adequate staining requires only 2–10 minutes of incubation. Actazolamide inhibits the staining reaction in specific concentrations. Actazolamidein vivo, 20 mg/kgi.v. to mice 30 minutes before sacrifice also inhibited the staining. The proportion phosphorous in the specific precipitate increases with KH2PO4 of the medium (shown by the addition of60Co and32P). An explanation of the reaction mechanism is given, based on the catalyzed loss of CO2 in the surface layer. The inclusion of phosphate in the medium makes this modification ofHäusler's method so sensitive that it shows carbonic anhydrase activity in for instance stratum spinosum of the skin.


Cobalt Surface Layer H2SO4 Reaction Mechanism KH2PO4 
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© Springer-Verlag 1967

Authors and Affiliations

  • Holger P. J. Hansson
    • 1
  1. 1.Departments of Pharmacology and DermatologyUniversity of UppsalaUppsalaSweden

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