Development of a high capacity radioimmunoassay procedure: ovine follicle stimulating hormone determination in blood plasma as a model.
A sensitive, specific and accurate homologous radioimmunoassay (RIA) for ovine follicle stimulating hormone (oFSH) has been developed, using a [125I]oFSH tracer and a polyclonal rabbit anti-OFSH-serum at a final dilution of 1:224,000. The separation of free and antibody-bound tracer is based on the double antibody solid phase system. The assay was found to be specific for oFSH; cross-reactivity with oLH, oPrl and oGH was lower than 0.2%. The detection limit was 7 pg per tube. Inter- and intra- assay coefficients of variation (CV) were 3.7-8.5% and 6.6%, respectively. The use of a few selfmade appliances in combination with a well-considered time-schedule enabled the processing of three thousand blood plasma samples in triplicate within two weeks. The particular advantage of the method described here is the fast, easy and safe separation of free and antibody-bound tracer with minimal handling of radioactive tubes.
Erkens
J H
JH
DLO-Research Institute for Animal Production, Schoonoord, Zeist, The Netherlands.
Visscher
A H
AH
Engel
B
B
D000818:Animals; D005069:Evaluation Studies as Topic; D005640:Follicle Stimulating Hormone / Q000097:blood* / Q000592:standards; D008954:Models, Biological; D011863:Radioimmunoassay / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012756:Sheep
D005640:Follicle Stimulating Hormone