A radioimmunoassay for thymosin alpha-1.
A new radioimmunoassay (RIA) is described for the quantitation of thymosin alpha-1 (alpha-1). The assay employs an antiserum specific for the COOH-terminal half segment 15-28 of alpha-1, synthetic alpha-1-(15-28) as the hormone standard, and a radioiodinated N alpha-acetyltryrosyl-alpha-1-(15-28) as the tracer. Since alpha-1-(1-28) lacks a phenolic ring for direct radioiodination, the N alpha-acetyltyrosyl-alpha-1-(15-28) was synthesized by the solid-phase method. The peptide bears a Tyr in place of Lys in position 14 of the natural peptide. It showed full alpha-1-(15-28) immunoreactivity and its radioiodinated derivative served as tracer in the RIA. An anti-alpha-1-(15-28) antiserum was raised in a rabbit and was shown to recognize alpha-1-(15-28) or its tyrosyl analogue, and the peptide, alpha-1-(1-28). But it did not recognize other thymic hormones or the biologically active segment 32-36 of thymopoietin, or structurally unrelated peptides. It could also detect natural alpha-1 cross-reacting material in the cytoplasm of cultured human thymic epithelial cells as measured by indirect immunofluorescence. In the RIA, as little as 9 pg of alpha-1-(15-28) equivalents in a 50 microliter sample could be detected. In addition, alpha-1-(1-28)-like immunoreactivity was quantitated in 6 human thymus homogenates and ranged from 0.5 to 4.5 ng/mg of protein.
G S
GS
Incefy
K
K
Ishimura
J G
JG
Wang
C G
CG
Unson
B W
BW
Erickson
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D006728:Hormones / Q000276:immunology; D006801:Humans; D011863:Radioimmunoassay; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D013997:Time Factors
D006728:Hormones; D013947:Thymosin; D000077596:Thymalfasin