Validation of a heterologous radioimmunoassay for insulin-like growth factor-I in bovine serum.
A quantitative, repeatable, heterologous radioimmunoassay (RIA) for insulin-like growth factor-I (IGF-I) was developed for bovine serum. Untreated serum could not be assayed due to interference by IGF-I-binding protein. Serum acidified to pH 3.6 in glycine-HCl (0.1 mol/l) for 24 h at 37 degrees C and neutralized with NaOH produced inhibition curves non-parallel to the [Thr59]-IGF-I standard. Neutralization by 40-fold dilution of acidified serum samples with assay buffer produced inhibition curves nearly parallel to the IGF-I standard. Complete parallelism was achieved by utilizing preprecipitated normal rabbit serum-sheep anti-rabbit gamma-globulin to separate antibody-bound 125I-labelled IGF-I from free 125I-labelled IGF-I. Recovery of IGF-I (1.3-52.3 fmol) added to serum was quantitative. The sensitivity of the RIA (n = 6) was 8.25 +/- 0.17 (S.E.M.) fmol. Intra- and interassay coefficients of variation were 3.03 and 4.95% respectively. Serum IGF-I levels measured in beef calves at weaning were positively correlated with weaning weight, total weight gain from birth to weaning and average daily weight gain. In conclusion, a heterologous RIA for IGF-I in beef serum which is sensitive, accurate, precise and repeatable has been developed.
Holland
M D
MD
Department of Animal Sciences, Colorado State University, Fort Collins 80523.
Hossner
K L
KL
Niswender
G D
GD
Elsasser
T H
TH
Odde
K G
KG
D000818:Animals; D002417:Cattle / Q000097:blood*; D005260:Female; D007334:Insulin-Like Growth Factor I / Q000097:blood*; D011863:Radioimmunoassay / Q000379:methods*; D013002:Somatomedins / Q000097:blood*
D013002:Somatomedins; D007334:Insulin-Like Growth Factor I