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Quantification of thromboxane B2

MC Michele Dei Cas
JR Jessica Rizzo
MS Mariangela Scavone
EF Eti Femia
GP Gian Marco Podda
EB Elena Bossi
MB Monica Bignotto
SC Sabrina Caberlon
MC Marco Cattaneo
RP Rita Paroni
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For TxB2 quantification, 3 mL of blood were collected in tubes without anticoagulants (in order to obtain serum). Non-anticoagulated blood was allowed to clot in a water bath at 37 °C for 1 h. Then centrifuged at 1400g for 15 min, and serum samples were frozen at − 20 °C until analysis.

TxB2, the stable metabolite of TxA2, was measured by an enzyme immunoassay. Frozen samples were thawed at 37 °C and opportunely diluted (between 1:2 and 1:750) with PBS and tested in duplicate. Samples were assayed in parallel with the standard calibration curve (detection limit = 1.6 pg/mL), prepared as outlined in the manufacturer's instruction. The 96-well plate was read at 450 nm wavelength using Ensight multimode Reader. Results were expressed as ng/mL.

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