New synthesis of immunogenic N6-isopent-2-enyladenosine-protein conjugate. Preparation, purification, and specificity of related antibodies.
An original procedure which preserves the structure of the sugar ring is described to link a plant hormone as N6-isopent-2-enyladenosine [( 9R]iP) onto a protein carrier to prepare a more specific immunogen. This cytokinin is bound to bovine serum albumin (BSA) and ovalbumin by a five-step procedure. These [9R]iP-protein conjugates have a maximal absorption at 269 nm and show molar ratios of hormone bound to proteins in the range of 12:1 and 18:1 for BSA and ovalbumin, respectively. Polyclonal antibodies were raised in rabbits against [9R]iP-BSA and were purified by affinity chromatography. Titers and specificity of the antisera and purified antibodies were determined by ELISA and RIA. These antibodies are highly specific for [9R]iP and do not cross-react with zeatin and ribosylzeatin. An immunoaffinity matrix was prepared with a capacity of 1 microgram of [9R]iP/mL of gel.
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Département de Biochimie des Glycoconjugués et Lectines Endogènes C.N.R.S., Orléans, France.
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Doumas
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Delmotte
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D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D003429:Cross Reactions; D003583:Cytokinins / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D007541:Isopentenyladenosine / Q000737:chemistry / Q000276:immunology*; D010047:Ovalbumin / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology*; D013056:Spectrophotometry, Ultraviolet
D000906:Antibodies; D003583:Cytokinins; D012710:Serum Albumin, Bovine; D007541:Isopentenyladenosine; D010047:Ovalbumin