Monoclonal antibodies to plant growth regulators. II. Indole-3-acetic acid

Mertens, Ralf; Eberle, J.; Arnscheidt, Angelika; Ledebur, A.; Weiler, Elmar W.

doi:10.1007/bf00401177

Cited by 54 publications

(32 citation statements)

References 8 publications

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“…As shown in Figure 3 such deviations could not be observed in the test extract. Together with the data shown in Figure 2, the results show that combination of separation techniques with ELISA (17) and IAA (18). In these studies, it became clear that the selection of mAB with specificities rivaling or even surpassing those of antisera is possible.…”

Section: -(2-ethylhexylamino)-2-methylpyrollo-[23-d]-pyrimidine and supporting

confidence: 57%

“…In these studies, it became clear that the selection of mAB with specificities rivaling or even surpassing those of antisera is possible. It was furthermore shown that, by employing appropriate screening protocols, the selection of high affinity mAB is possible (17,18). Both specificity and affinity need to be maximized in order to allow for optimum immunoassay performance.…”

Section: -(2-ethylhexylamino)-2-methylpyrollo-[23-d]-pyrimidine and mentioning

confidence: 99%

“…Due to their extreme usefulness as standardizable reagents (22) monoclonal antibodies specific for plant growth regulators are being developed (17,18). In this paper we report on the production of mAB with high affinity and specificity for ZR and DHZR, their free bases and phosphate esters.…”

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confidence: 99%

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Monoclonal Antibodies to Plant Growth Regulators

Eberle¹,

Arnscheidt²,

Klix³

et al. 1986

Plant Physiol.

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Section: -(2-ethylhexylamino)-2-methylpyrollo-[23-d]-pyrimidine and supporting

confidence: 57%

Section: -(2-ethylhexylamino)-2-methylpyrollo-[23-d]-pyrimidine and mentioning

confidence: 99%

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Monoclonal Antibodies to Plant Growth Regulators

Eberle¹,

Arnscheidt²,

Klix³

et al. 1986

Plant Physiol.

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“…To raise an antibody against IAA, it must be coupled to a carrier protein. The nitrogen in the indole ring (IAA-N), [9][10][11] the carboxyl group (IAA-C), 12,13) and the hetero atoms introduced on a benzene ring of IAA (IAA-H) 14,15) are often used for the conjugation. The antibodies raised against IAA-C showed higher and wider cross-reactivity to IAA conjugates than the antibodies against IAA-N or IAA-H, and required methylation of samples for the analysis of free IAA in plants.…”

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confidence: 99%

Expression of a Single-chain Antibody against Indole-3-acetic Acid inEscherichia coli

Mitani

Matsumoto

2004

Bioscience, Biotechnology, and Biochemistry

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A hybridoma cell line that produces a monoclonal antibody specific for indole-3-acetic acid (IAA) was prepared. The DNA fragments coding the variable regions of the light and the heavy chains of the antibody were prepared by PCR using the cDNA of the antibody as a template. A chimera DNA for a single chain variable fragment (scFv) was constructed, and expressed in Escherichia coli. The scFv antibody expressed in E. coli as well as the original monoclonal antibody showed a specific binding to IAA.Key words: single chain variable fragment (scFv); indole-3-acetic acid (IAA); enzyme-linked immunosorbent assay (ELISA)

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“…We report here the development of an indirect competitive ELISA for the quantitation of extensin precursors. This type of ELISA has been demonstrated to be useful and sensitive in the assay of important biological molecules (10). ' Deglycosylation of P1 and P2 involved HF solvolysis of the saccharide component using anhydrous HF with 10% methanol as the scavenger for 1 h at 0°C (12).…”

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confidence: 99%

Detection of Glycosylated and Deglycosylated Extensin Precursors by Indirect Competitive ELISA

Conrad

Lamport²,

Hammerschmidt³

1987

Plant Physiol.

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A competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for the rapid quantitation of the glycosylated and deglycosylated forms of the monomeric soluble extensin precursor subunits Pl and P2. A log-linear response range for each kind of precursor in the competition curve was between 0.01 and 100 nanograms per milliliter.Extensin is a plant cell wall-bound hydroxyproline-rich glycoprotein. Two different soluble forms of extensin were recently isolated from salt eluates of intact tomato cell suspension cultures (12). These HRGPs3 were shown to be different in amino acid composition. These two soluble extensins, P1 and P2, are monomeric precursors to insoluble network extensin. Although the evidence is correlative, extensin may function as part of the resistance mechanism against certain biological stresses. Extensin accumulates in cell walls in response to stress stimuli such as wounding (14), pathogen attack (2,3,8,9), and heat shock (13), but the precise role of this response is not known. Key to understanding the role ofextensin in these responses is the ability to quantitate the amounts of extensin that accumulates in response to stress. Satisfactory evidence has not been provided to date, partly because ofthe inability to quantitate soluble extensin accurately in plant cell walls.Current techniques for quantitation of the accumulation of HRGPs in cell walls involve spectrophotometric measurement of hydroxyproline released after acid hydrolysis (7). Such methods are, however, slow, insensitive, and nonspecific. Immunological methods, such as ELISA are rapid and highly sensitive (11). We report here the development of an indirect competitive ELISA for the quantitation of extensin precursors. This type of ELISA has been demonstrated to be useful and sensitive in the assay of important biological molecules (10). Deglycosylation of P1 and P2 involved HF solvolysis of the saccharide component using anhydrous HF with 10% methanol as the scavenger for 1 h at 0°C (12).Rabbit Immunization. P1, P2, and deglycosylated P1 and P2 (DPl and DP2) were used as immunogens. Initial injection of all four immunogens employed a modification of the multiple site method of Vaitukaitis et al. (15). For the two glycosylated immunogens, P1 and P2, 0.5 mg precursor in 1.0 ml of 0.9% saline was emulsified with 2.0 ml Freund's complete adjuvant. For the two deglycosylated immunogens DPl and DP2, 0.5 mg precursor in 0.5 ml of 0.9% saline was emulsified with 1.5 ml Freund's complete adjuvant. With all four precursors, the preparation was injected intradermally into four sites on the back of a New Zealand white doe rabbit. Subsequent injections were made at 6 week intervals using Freund's incomplete adjuvant emulsified in the same ratios and concentrations as described above, but at one-half the volume. Rabbits were bled through the marginal ear vein and sera purified by three (NH4)2, S04 (35% saturated) precipitations (4) followed by dialysis overnight at 4°C against 0.1 M phosphate buffer in 0.15 M saline (...

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Monoclonal antibodies to plant growth regulators. II. Indole-3-acetic acid

Cited by 54 publications

References 8 publications

Monoclonal Antibodies to Plant Growth Regulators

Monoclonal Antibodies to Plant Growth Regulators

Expression of a Single-chain Antibody against Indole-3-acetic Acid inEscherichia coli

Detection of Glycosylated and Deglycosylated Extensin Precursors by Indirect Competitive ELISA

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