Validation of an enzime linked immunosorbent assay for the detection of progesterone in Santa Inês ewes / Validação de um ensaio imunoenzimático para detecção de progesterona em ovelhas Santa Inês
de Miranda e Silva Chaves, Camila; Cavallaro Machado, Débora; Lúcia Gonçalves Garcia Gazzetta, Sylvia; Maria Roncato Duarte, Keila; Lopes Dias da Costa, Ricardo.
Bol. ind. anim. (Impr.);
71: 52-52, 2014.
Artigo
em Inglês
|
LILACS-Express
| ID: biblio-1466629
Resumo
The success of reproductive management in the sheep industry depends on several factors. Among them, early pregnancy diagnosis stands to improve the rates of exploration of sheep livestock, allowing the identification of fertility problems, disposal of infertile animals, food supplementation of pregnant females and maximizing production and profits. Therefore, this work aimed to develop an Enzyme Linked Immunosorbent Assay (ELISA) for detection of progesterone in peripheral blood samples from sheep of Santa Inês breed. A calibration curve was made on the test plate at which a fraction of 100 L of diluted antigen was deposited in the wells. The antigen (P4) was diluted sample in methanol/water and incubated at 37ºC for one hour; then 200 L were deposited from the PBS-BSA 1% and was followed by incubation overnight at 10ºC. 100 L of anti-P4 serum produced in rabbits were added and another incubation at 37ºC for one hour was performed, followed by two washes with PBS-T-G. Then it was added 100 L of the conjugate anti rabbit peroxidase labeled, followed by another incubation at 37ºC for one hour; two washing were done with PBS-T-G and drying the plate; revelation with TMB (Sure Blue) was performed using 100 L of Sure Blue for 10 minutes and the reaction stopped, the plate was read at 595 nm; another revelation was done with PNPP and the plate reading was done at 450 nm. The rO artigo não possui resumo em português.
Biblioteca responsável:
BR68.1
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