The Development of an Indirect Enzyme Linked Immunoassay for Abscisic Acid

Ross, G. J.; Elder, Peter A.; McWha, James A.; Pearce, David; Pharis, Richard P.

doi:10.1104/pp.85.1.46

Cited by 44 publications

(19 citation statements)

References 12 publications

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“…Hundred millilitre samples (extra cellular culture filtrates), containing 1 mg of Butylated Hydroxy Toluene (BHT) to prevent oxidation of the hormones (Ross et al 1987), were centrifuged at 959 g for 10 min then the samples were extracted. Extraction, purification and quantitative determination of IAA, GA 3 , zeatin and ABA were carried out, with minor modifications, according to the methods of Ü nyayar et al (1996) and Baydar & Ü lger (1997).…”

Section: Methodsmentioning

confidence: 99%

Auxin, Gibberellin, Cytokinin and Abscisic Acid Production in Some Bacteria

Karadeniz

Topcuoglu

Inan

2006

World J Microbiol Biotechnol

177

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In this study, auxin (indole-3-acetic acid), gibberellin, cytokinin (zeatin) and abscisic acid production were investigated in the culture medium of the bacteria Proteus mirabilis, P. vulgaris, Klebsiella pneumoniae, Bacillus megaterium, B. cereus, Escherichia coli. To determine the levels of these plant growth regulators, high performance liquid chromatography (HPLC) technique was used. Our findings show that the bacteria used in this study synthesized the plant growth regulators, auxin, gibberellin, cytokinin and abscisic acid.

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Section: Methodsmentioning

confidence: 99%

Auxin, Gibberellin, Cytokinin and Abscisic Acid Production in Some Bacteria

Karadeniz

Topcuoglu

Inan

2006

World J Microbiol Biotechnol

177

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“…The samples were redissolved in 10 pL of 100% methanol and methylated with 500 pL of diazomethane at room temperature. The samples were then analyzed by GC-MS-selected ion monitoring using the procedure of Ross et al (1987).…”

Section: Aba Extraction and Gc-ms Analysismentioning

confidence: 99%

Competitive Inhibition of Abscisic Acid-Regulated Gene Expression by Stereoisomeric Acetylenic Analogs of Abscisic Acid

et al. 1993

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S7N OW9 (S.R. A.) l h e properties of two enantiomeric synthetic acetylenic abscisic acid (ABA) analogs (PBI-51 and PBI-63) in relation to ABA-sensitive gene expression are reported. Using microspore-derived embryos of Brassica napus as the biological material and their responsiveness to ABA in the expression of genes encoding storage proteins as a quantitative bioassay, we measured the biological activity of PBI-51 and PBI-63. Assays to evaluate agonistic activity of either compound applied individually showed a dose-dependent increase in napin gene expression on application of PBI-63. Maximal activity of about 40 p~ indicated that PBI-63 was an agonist, although somewhat weaker than ABA. PBI-63 has a similar stereochemistry to natural ABA at the junction of the ring and side chain. In contrast, PBI-51 showed no agonistic effects until applied at 40 to 50 p~.Even then, the response was fairly weak. PBI-51 has the opposite stereochemistry to natural ABA at the junction of the ring and side chain. When applied concurrently with ABA, PBI-63 and PBI-51 had distinctly different properties. PBI-63 (40 p~) and ABA (5 PM) combined gave results similar to the application of either compound separately with high levels of induction of napin expression. PBI-51 displayed a reversible antagonistic effect with ABA, shifting the typical ABA dose-response curve by a factor of 4 to 5. This antagonism was noted for the expression of two ABA-sensitive genes, napin and oleosin. To test whether this antagonism was at the leve1 of ABA recognition or uptake, ABA uptake was monitored in the presence of PBI-51 or PBI-63. Neither compound decreased ABA uptake. Treatments with either PBI-51 or PBI-63 showed an effect on endogenous ABA pools by permitting increases of 5-to 7-fold. It is hypothesized that this increase occurs because of competition for ABA catabolic enzymes by both compounds. l h e fact that ABA pools did not decrease i n the presence of PBI-51 suggests that PBI-51 must exert its antagonistic properties through direct competition with ABA at a hormone-recognition site.

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“…This included generations of a new ABA-mAb and development of an indirect ELISA [20], which provides substantial amplification of the response. To our knowledge, this combination of mAb with an indirect ELISA has been applied only few times before for ABA quantitation [2,9,19,221, while all other studies dealing with immunoassays for ABA used polyclonal Ab [4,8,14,24,251 or mAb coupled with RIA [15,18,211 rather than with ELISA. Employing these immunological techniques minimized the need for purification of the ABA extracts, so that a large number of samples could be processed with little procedural effort, and correspondingly little sample loss.…”

Section: Discussionmentioning

confidence: 99%

“…Unlike previous ELISA quantitations of ABA, which coupled the hormone with alkaline phosphatase [4,9,251, this report describes an indirect ELISA method, that provides substantial amplification of the response by employing a second antibody, goat anti-mouse, coupled to the enzyme horse-radish peroxidase (HRP) [20]. A similar approach is now used by several other groups [19,2,221. Regarding previous investigations dealing with rice leaf senescence [3,10, …”

Section: Introductionmentioning

confidence: 99%

Characterization and use in ELISA of a new monoclonal antibody for quantitation of abscisic acid in senescing rice leaves

1993

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A new monoclonal antibody (mAb) was generated against abscisic acid (ABA), and an indirect enzymelinked immunosorbent assay (ELISA) using this mAb was developed for convenient quantitative analysis of ABA levels in rice leaf extracts. The mAb, raised against (+)-ABA conjugated to bovine serum albumin (BSA) through its carboxyl group (C,), reacted preferentially with the (+)-ABA enantiomer, and equally well with both free and methyl-ester (+)-ABA. Cross-reactivity with several ABA-related compounds was negligible. Linearity was obtained between 3 and 1OOOpmol of (+)-ABA. The ABA-mAb was further used to quantitate pmol quantities of (+)-ABA in attached and detached rice leaves. Results obtained with such ELISA quantitation showed an increase in the free ABA content of detached rice leaves at progressive stages of senescence, which was regarded as a senescence-related response. This quantitation compared favorably with other presently used techniques for ABA determination, with regard to their detection limits, cost and assay time. The results suggest that the combination of a specific mAb with a sensitive ELISA technique is quite promising for quantitation of ABA.

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The Development of an Indirect Enzyme Linked Immunoassay for Abscisic Acid

Cited by 44 publications

References 12 publications

Auxin, Gibberellin, Cytokinin and Abscisic Acid Production in Some Bacteria

Auxin, Gibberellin, Cytokinin and Abscisic Acid Production in Some Bacteria

Competitive Inhibition of Abscisic Acid-Regulated Gene Expression by Stereoisomeric Acetylenic Analogs of Abscisic Acid

Characterization and use in ELISA of a new monoclonal antibody for quantitation of abscisic acid in senescing rice leaves

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