Determination of histamine secretion by human mast cells


Histamine is a mediator involved in inflammatory and allergic phenomena with immediate manifestation. It is massively secreted by mast cells and basophilic polynuclei after contact with an allergen.

The determination of histamine is therefore a relevant test for the identification of potential allergens.


Two compatible fluorophores, donor (europium cryptate) and acceptor (d2), are bound to an anti-histamine antibody and to histamine respectively.

Histamine from the sample competes with histamine linked to the acceptor fluorophore for the binding to the antibody, preventing the time-resolved energy transfer (TR-FRET) between the 2 fluorophores.


Format96 wells 
Cellular modelHuman Mast Cells-1 
StimulationA23187 / PMA      
Reference moleculeQuercetol  
Incubation18H at 37°C under 5% CO
RevelationHTRF ASSAY
Format384 wells
DetectionDetection Excitation wavelength : 337 nm
Emission wavelength 1: 615 nm (donor fluorophore emission)
Emission wavelength 2: 665 nm (acceptor fluorophore emission)

Notes :

  • Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration)
  • Cell viability is assessed in parallel

Recommendation :

  • Ensure solubility of sample in a solvent compatible with cell culture

Histamine secretion in the presence of A23187 and PMA

Inhibition of histamine secretion in the presence of Quercetol