Prostaglandins (PGs) and their metabolites are measured by gas chromatography with mass spectrometric or electron capture detection, high-pressure liquid chromatography (HPLC) with ultraviolet or fluorescent detection, or radioimmunoassay. Of these methods, radioimmunoassay offers the advantages of high sensitivity, specificity, simplicity, and economy. This chapter presents a method that is applied to the measurement of PGs, PG metabolites, thromboxanes, and other eicosanoids by radioimmunoassay. It discusses the preparation of immunogenes. The PG is coupled to porcine thyroglobulin by the mixed anhydride method. Other proteins as bovine serum albumin, bovine γ globulin, or keyhole limpet hemocyanin may be used. Immunization is carried out in rabbits or chickens. Rabbits are injected with this emulsion in multiple (30–40) subcutaneous sites on the back using the same syringe and needle. Chickens are injected in the wing web, back muscles, and breast muscles. To be assured of obtaining antibodies, it is recommended that two to four animals be injected with the same immunogen. The chapter presents the characterization of antisera.
ASJC Scopus subject areas
- Molecular Biology