DUBOVÁ, Jaroslava, Julie Olegivna ROTKOVSKÁ, Alena REKOVÁ and Břetislav BRZOBOHATÝ. Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene. In Progress in Basic, Applied and Diagnostic Histochemistry. Brno: Masarykova univerzita v Brně, 2002, p. 15. ISBN 80-210-2998-6.
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Basic information
Original name Optimisation of Immunohistochemical Detection of Cytokinins in Transgenic Arabidopsis Seedlings Expressing Activated ipt, a Cytokinin Biosynthesis Gene
Name in Czech Optimalizace imunohistochemické detekce cytokininů u transgenních klíčních rostlin Arabidopsis exprimujících ipt, gen pro biosyntézu cytokininů
Authors DUBOVÁ, Jaroslava (203 Czech Republic), Julie Olegivna ROTKOVSKÁ (804 Ukraine), Alena REKOVÁ (203 Czech Republic) and Břetislav BRZOBOHATÝ (203 Czech Republic, guarantor).
Edition Brno, Progress in Basic, Applied and Diagnostic Histochemistry, p. 15-15, 2002.
Publisher Masarykova univerzita v Brně
Other information
Original language English
Type of outcome Proceedings paper
Field of Study 10600 1.6 Biological sciences
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14310/02:00006761
Organization unit Faculty of Science
ISBN 80-210-2998-6
Keywords in English cytokinins; immunohistochemistry; Arabidopsis
Tags Arabidopsis, cytokinins, Immunohistochemistry
Changed by Changed by: RNDr. Jaroslava Dubová, CSc., učo 1244. Changed: 7/3/2006 18:57.
Abstract
Aseptically grown Arabidopsis seedlings were collected, fixed in PFA - GA mixture. Indirect immunohistochemistry on cryostat sections was used. Primary antibodies against zeatin-riboside and isopentenyladenine-riboside were used respectively. Goat anti-rabbit IgG conjugated with AP was used as a secondary antibody. AP activity was detected by reaction with BCIP and NBT. The best results of immunostaining were obtained by higher dilution of primary antibodies and lowering the concentration of BCIP and NBT. Arabidopsis seedlings with activated ipt showed higher the amount of reaction product in comparison with the wild type plants. Accumulation of cytokinins was established immunohistochemically during culture period in transgenic Arabidopsis seedlings, which is in accordance with the biochemical analyses. Modified immunohistochemical detection of free bases of cytokinins can be used for the semi quantitative evaluation of cytokinin content in Arabidopsis seedlings.
Links
IAA5004001, research and development projectName: Systém aktivace transkripce pro studium vztahu mezi metabolizmem a účinkem cytokininů u Arabidopsis a tabáku
Investor: Academy of Sciences of the Czech Republic, Transkription activation system for studying the relationship between cytokinin metabolism and action in Arabidopsis and tobacco
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