Development and application of 5-HETE radioimmunoassay (RIA)

Conference · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5126312

Hwang, D H

5-hydroxyeicosatetraenoic acid (5-HETE) lactone was converted to omega-aminopropylamide and coupled to BSA using 1,5-difluoro-2,4-dinitrobenzene as a coupling agent. Antibodies were developed by immunizing rabbits with 5-HETE-BSA conjugate for 11 months with monthly boosters. The antiplasma at 1 to 800 dilution bound 35% of the total radioactivity of the tritiated 5-HETE (150 Ci/mmole). The detection limit of the RIA (one SE of the buffer control) was 19 picogram. The antiplasma showed the greatest cross reactivity (0.24%) with LTB/sub 4/ among compounds tested (arachidonic acid, diarachidonyl phosphatidyl choline, linoleic acid, dilinoeyl phosphatidyl choline, linolenic acid, LTB/sub 4/, 5,12-diHETE, 5,15-diHETE, 15-HETE, 12-HETE, TXB/sub 2/ and other major prostaglandins). When the ethanol extract of rat macrophages (peritoneal), stimulated by Calcium ionophore, was injected into HPLC and each fraction was assayed for 5-HETE by RIA, there was only one immunoreactive peak which appeared at fractions corresponding to the elution time of 5-HETE standard. RIA for LTB/sub 4/ showed that the sample also contained LTB/sub 4/ and 12-HETE. Thus, this antiplasma can be used to monitor 5-HETE, which is a relatively stable relic of arachidonic acid metabolism via 5-lipoxygenase pathway, synthesized from the endogenous precursor in various biological samples.

• Research Organization: Louisiana State Univ., Baton Rouge
• OSTI ID: 5126312
• Report Number(s): CONF-8606151-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
ARACHIDONIC ACID
CARBOXYLIC ACIDS
EICOSANOIC ACID
ENZYMES
ESTERS
HETEROCYCLIC COMPOUNDS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LACTONES
MAMMALS
METABOLITES
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
OXYGENASES
RABBITS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES