A radioimmunoassay for 6-keto-prostaglandin F1 alpha utilizing an antiserum against 6-methoxime-prostaglandin F1 alpha. Application to study renal in vitro synthesis of prostacyclin

Prostaglandins
E Oliw

Abstract

PGI2 and 6-keto-PGF1 alpha were converted to 6-methoxime-PGF1 alpha (6-MeON-PGF1 alpha) by treatment with methoxyamine HCl in acetate buffer. The formed 6-MeON-PGF1 alpha was measured by radioimmunoassay. Antisera were raised in rabbits after immunization against 6-MeON-PGF1 alpha-BSA conjugate. Diluted 1:20.000 to bind 50% of the tracer (3H-6-MeON-PGF1 alpha, 100 Ci/mmol), the antiserum cross reacted 0.8% with PGE2, 1% with PGF2 alpha and less than 0.2% with PGD2, PGF1 alpha, PGF2 beta and TXB2. The radioimmunoassay was used to estimate release of PGI2 and 6-keto-PGF1 alpha from chopped rabbit renal medulla and cortex incubated in Krebs-Ringer bicarbonate buffer (37 degrees C, 30 min). The 6-keto-PGF1 alpha radioimmunoassay was validated in biological samples by mass fragmentography. The chopped medulla (n = 5) released 38-+9 ng/g/min and the cortex (n = 5) 4.7-+2-0 ng/g/min, while the release of immunoreactive PGE2 (iPGE2) and iPGF2 alpha was 171-+26 and 74-+13 ng/g/min from the medulla and 4.3-+1.3 and 2.7-+0.3 ng/g/min from the cortex, respectively. The results confirm previous findings, which indicate that in the renal medulla prostaglandin endoperoxides are mainly transformed to prostaglandins, while in the cortex transfo...Continue Reading

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