A new method for detecting histamine release
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Glass microfibres have been found to bind histamine with high affinity and selectivity. A new test for measuring basophil hisamine release has been developed using the glass microfibres as a solid phase. Glass microfibres are crushed and fixed to the bottom of microtitre plates with a water-soluble glue. Histamine release is performed in the glass microfibre-prepared microtitre plates by challenging 100 μl washed blood with 20 μl antigen per well for 90 min at 37°C. Released histamine is bound with high affinity to the glass microfibres, since 90% of histamine in the solution is adsorbed to the fibres. After incubation the microtitre plate is washed with H2O to remove cells and interfering substances. Fibre-bound histamine is detected by the fluorometrico-phthaldialdehyde method. The sensitivity of the assay is 0.63 ng histamine, 2 HCl and the histamine standard curve is linear up to at least 5 ng histamine, 2 HCl. Optimal conditions for the new assay are described. After challenge with anti-Ige a comparison with the conventional histamine release from Ficoll-Hypaque-isolated leukocytes showed almost identical results.
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- P. Stahl Skov andS. Norn,A simplified method for measuring basophil histamine release and blocking antibodies in hay fever patients. Basophil histamine content and cell preservation, Acta allerg.32, 170–182 (1977).Google Scholar