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We have established a double antibody RIA using a rabbit antiserum prepared against reduced, carboxymethylated (RCXM) human LH alpha-subunit, with RCXM-alpha as tracer and standard. This antiserum did not cross-react with any native gonadotropins or subunit, and reacted only weakly with RCXM-alpha. A tryptic digest of RCXM alpha-subunit was completely reactive, while chymotryptic digestion abolished all immunoreactivity. By testing with separate tryptic fragments, the recognition site could be localized to a segment close to the amino-terminus of the peptide chain. When applied to measurement of serum and urine, an immunoreactive species, parallel to RCXM alpha-subunit by serial dilution, was found in concentrations of 1-2 ng/ml in serum and 3-4 ng/ml in urine. Similar levels of the immunoreactive component were found in conditions of elevated gonadotropins (e.g. pregnancy) as well as gonadotropin deficiency(panhypopituitarism and Kallmann's syndrome). After stimulation with LHRH, no rise was noted at times up to 6 h despite the fact that both LH and LH-alpha were elevated. The data indicate that the sequence-specific antiserum may be detecting an immunoreactive form of alpha-subunit of LH whose kinetics of appearance and disappearance differs from those of the native subunit.
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
April 1975,
Acta endocrinologica,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
April 1966,
Metabolism: clinical and experimental,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
January 1967,
Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.),
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
January 1978,
Endocrine research communications,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
September 1976,
General and comparative endocrinology,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
July 1975,
The Journal of clinical endocrinology and metabolism,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
July 1970,
Research communications in chemical pathology and pharmacology,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
July 1974,
The Journal of laboratory and clinical medicine,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
July 1966,
Endocrinology,
H T Keutmann, and
I Z Beitins, and
L Johnson, and
J W McArthur
June 1976,
Analytical biochemistry,
