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Detection and determination of Locusta migratoria trypsin by radioimmunoassay
Author(s) -
Sakal Edna,
Birk Yehudith,
Applebaum Shalom W.
Publication year - 1992
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.940200207
Subject(s) - locust , antiserum , trypsin , polyclonal antibodies , biology , radioimmunoassay , hemolymph , biochemistry , migratory locust , heterologous , microbiology and biotechnology , midgut , enzyme , western blot , chymotrypsin , antibody , immunology , larva , botany , gene
Polyclonal antibodies were produced against trypsin‐like enzyme TLE‐2, one of two trypsins isolated from the midgut cecae of the locust Locusta migratoria . Using those antibodies a heterologous RIA was developed. The specificity of the antibodies was demonstrated by immunoblot technique and cross‐reactivity experiments. Positive blot reaction with the antiserum was observed with locust trypsins and binding competition was obtained only with the second trypsin from the locust, TLE‐1. Trypsins from bovine and hog as well as trypsins from Tenebrio molitor and Tribolium castaneum and locust chymotrypsin did not inhibit TLE‐2 binding to the antiserum even at concentrations 1,000‐fold greater than that of locust TLE‐2. The concentrations of TLE‐2 in cecal fluid, cecal wall, hemolymph, and fat‐body were 2,790, 60.4, 6.4, and 0.7 ng per mg tissue, respectively. © 1992 Wiley‐Liss, Inc.