Produção de anticorpos IgY de galinhas e IgG de coelhos para análise de auxina e citocininas. Seropédica: UFRRJ, 2008. 61 p. / Production of antibodies IgY from chicken and IgG from rabbits for analysis of auxin and cytokinins. Seropédica: UFRRJ, 2008. 61 p

AUTOR(ES)
DATA DE PUBLICAÇÃO

2008

RESUMO

The determination of plant hormones can be an excellent tool for to study the plant development. Today, the complexity of methods, equipment and reagents of high cost limit the use with practice of routine. Some advantages of methods immunoassay can exceed those limitations, showing a potential of practical use in determination of plant hormones. The test Enzyme-Linked Immunosorbent Assay (ELISA) is utilized in detection of molecules with low molecular weight. However, the ELISA requires specific antibodies against molecules of interest, in the case, plant hormones. Today in the market there is not available antibodies for the determination of hormonal molecules. Faced with that, proposed be produced and characterize antibodies against molecules of AIA, 2ip and zeatin and subsequently, for use the detection those molecules. For that, the hormonal molecules were conjugated with protein (BSA) for subsequent immunization of chicken or rabbits. From the serum of rabbits or yolks eggs the antibodies were purified and characterized through the test of precipitation of double diffusion, SDS-PAGE and test ELISA. The antibodies that presented better results were utilized in detection of plant hormones molecules in samples of plantlets in vitro of gerbera. The test of precipitation of double diffusion revealed that the antibodies productized were capable of will detect the molecule in study. Through in the SDS-PAGE was verified that the antibodies obtained in eggs yolks of chicken presented superior purity than them obtained in serum of rabbits. From the results of the test ELISA, observed itself that the antibodies against AIA did not present potential of practical use. The detection of plant hormones in samples of tissue stayed restricted to 2ip and zeatin. The detection of those two molecules in crude extract, obtained from plantlets in vitro of gerbera, revealed that the level endogenous of zeatin was higher of level 2ip. Plantlets with six weeks after multiplication presented higher level of zeatin than plantlets with one week after multiplication. For the 2ip, that difference was not evident. The use of antibodies obtained in eggs yolks of chicken permitted the determination of zeatin and 2ip in samples plant utilizing the test ELISA.

ASSUNTO(S)

produção de anticorpos elisa determinação hormônios vegetais gérbera in vitro. fisiopatologia da reproducao animal key words: production of antibodies, elisa, determination of plant hormones, gerbera in vitro.

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