Characierization of monoclonal antibodies specific for isopentenyl adenosine derivatives occurring in transfer RNA

Woodsworth, Mary L.; Latimer, L.; Janzer, Janice J.; McLennan, B. D.; Lee, Jeremy S.

doi:10.1016/0006-291x(83)90851-3

Cited by 15 publications

(6 citation statements)

References 10 publications

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“…The specificities of the antibodies developed in this study (Tab. 2) were similar to those noted by Woodsworth et al (1983), Their radioimmunoassay indicated that certain of their antibodies had high specificity for iPA. This was in contrast to the monoclonal antibodies developed by Trione et al (1985) against ZR which cross-reacted with a variety of structurally similar cytokinins.…”

Section: Discussionsupporting

confidence: 79%

“…Monoclonal antibodies have been used in immunoassays for abscisic acid (ABA; Mertens et al 1983), iso-The use of immunoassay for detection and quantifica-pentenyl adenosine (iPA; Woodsworth et al 1983), zeation of plant growth regulators has gained widespread tin riboside (ZR; Trione et al 1985;Eberle et al 1986), use in plant physiology. Many of these assays have and dihydrozeatin riboside (diHZR; Eberle et al 1986).…”

Section: Introductionmentioning

confidence: 99%

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Monoclonal antibodies against the plant cytokinin isopentenyl adenosine

Trione

Krygier

Kathrein

et al. 1987

Physiologia Plantarum

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Sixteen monoclonal antibodies against isopentenyl adenosine (iPA) were developed and characterized for reactivity towards this cytokinin and structurally related molecules by use of a competition fluorescence enzyme immunoassay. Antibodies with suitable affinity and specificity were used in an immunoassay to detect and quantify isopentenyl adenosine. Logit/log plots of fluorescence vs pmol of the cytokinin indicated that the assay had a measurement range of 0.03–256 pmol (10–8500 pg) with high linearity (r =–0.98). This competition fluorescence enzyme immunoassay showed that three antibodies cross‐reacted with N‐6‐benzyladenine and its riboside: cross‐reactivity with dihydrozeatin and its riboside, cis‐zeatin, trans‐zeatin riboside, adenine and adenosine was minor. When an immunoaffinity‐HPLC technique was used to measure the cross‐reactivity of two of the antibodies in the presence of known quantities of multiple cytokinins, iPA was bound in preference to the other cytokinins. One of the antibodies was used to quantify this cytokinin in developing wheat (Triticum aestivum L. cv. Stephens) seeds and in wheat seeds spiked with known amounts of certain other cytokinins. The use of these antibodies in immunoassay in combination with HPCL for quantification of iPA in plant extracts is discussed.

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Section: Discussionsupporting

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Monoclonal antibodies against the plant cytokinin isopentenyl adenosine

Trione

Krygier

Kathrein

et al. 1987

Physiologia Plantarum

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“…The ability to prepare mixtures of mcABs may be very useful for specific removal of selected cytokinins from a plant extract for subsequent analysis on HPLC or similar instrumentation as suggested by MacDonald et al (1981). Alternatively, preparation of mcABs with high specificity, such as those described by Woodsworth et al (1983) may allow direct quantitation of cytokinins in plant extracts without the use of additional analytical techniques.…”

Section: Discussionmentioning

confidence: 97%

“…Woodsworth et al (1983) prepared mcABs against iPA; certain of these antibodies were highly specific for iPA while others cross-reacted with selected structurally related molecules. The enhanced cross-reactivity of mcABs as compared to polyclonal antiserum is well documented for other antigens (Lane and Hoeffler 1980, PiUemar and Weissman 1981, Dulbecco et al 1981) and possible explanations have been offered (Lane and Koprowski 1982).…”

Section: Discussionmentioning

confidence: 99%

The development of monoclonal antibodies against the cytokinin zeatin riboside

Trione

Krygier

Banowetz

et al. 1985

J Plant Growth Regul

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Abstract. Seven monoclonal anti-zeatin riboside antibodies were characterized by radioimmunoassay (RIA) and found to measure femtomole (10-15 M) quantities (-20 pg) of this cytokinin. The antibodies had different measuring ranges defined by the linear portion of the logit/log plots; slopes and intercepts of the line varied considerably between the antibodies. Competitive binding trials against cis-zeatin riboside (cZR), dihydrozeatin riboside (diHZR), zeatin (Z), and isopentenyl adenosine (iPA) showed differences among the seven antibodies in their cross-reactivities towards these structurally related cytokinins. It was possible to combine selected antibodies to provide a mixture with a predictable measuring range and cross-reactivity; the ability to prepare a highly specific reagent in this manner with well-defined reactivity was noted and differences between monoclonal antibody and polyclonal antiserum probes for measurement of cytokinins were discussed.Cytokinins play important roles in many plant physiological processes including the regulation of growth and development. Plant hormone research has been hindered by the lack of specific and sensitive methods for qualitative and quantitative assays, in part because cytokinins and other growth regulators

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“…An antibody with extensive cross-reactivity is desirable for use in immunoaffinity matrices because it provides the potential for simultaneous isolation and subsequent HPLC purification of a large number of different cytokinins (6). The anti-cis-[9R]Z F-ELISAs using these three antibodies had detection limits and measuring ranges similar to previously characterized anti-cytokinin antibodies (2)(3)(4)(5)(6)(7)(8)(9)(10). Although eis-[9R]Z has been detected in a range of other plant tissues (10)(11)(12)(13), no endogenous eis-[9R)Z was detected in the wheat leaves examined in these studies.…”

Section: Resultsmentioning

confidence: 73%

Monoclonal Antibodies Against the Plant Cytokinin, cis-Zeatin Riboside

Banowetz¹

1993

Hybridoma

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Three monoclonal antibodies (two IgG1 and one IgA) prepared against the plant cytokinin, cis-zeatin riboside, were characterized for use in a fluorescence competition enzyme linked immunosorbant assay (F-ELISA). Immunoassays that employed each antibody detected femtomole quantities of the homologous cytokinin and characteristic quantities of structurally-related cytokinins. One of the antibodies was used to quantify HPLC-purified cis-zeatin riboside recovered from spiked samples of wheat leaves. These are the first monoclonal antibodies against a cis- derivative of a plant cytokinin and will be useful for purification and quantification of cis-zeatin and the 9-riboside and 9-glucoside in plant samples.

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Characierization of monoclonal antibodies specific for isopentenyl adenosine derivatives occurring in transfer RNA

Cited by 15 publications

References 10 publications

Monoclonal antibodies against the plant cytokinin isopentenyl adenosine

Monoclonal antibodies against the plant cytokinin isopentenyl adenosine

The development of monoclonal antibodies against the cytokinin zeatin riboside

Monoclonal Antibodies Against the Plant Cytokinin, cis-Zeatin Riboside

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