[29] Enhanced chemiluminescent reactions catalyzed by horseradish peroxidase

Thorpe, G. H. G.; Kricka, Larry J.

doi:10.1016/0076-6879(86)33078-7

Cited by 228 publications

(119 citation statements)

References 30 publications

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“…The blots were incubated for 1 h at room temperature with 1% casein in TBS with 0.1% Tween 20 (TBST) and incubated with primary antibodies diluted in blocking buffer. The blots were washed twice for 10 min with TBST and twice with TBS for 10 min and then incubated with a 1:5000 dilution of horseradish peroxidase goat anti-mouse or horseradish peroxidase-protein A in TBST for 30 min at room temperature; the blots were washed as before, and the proteins were visualized by ECL (22). The anti-LRP1, anti-HA, anti-His, anti-phosphotyrosine, and anti-Myc antibodies were used at 1:100, 1:20, 0.1 g/ml, 1:10 and 1:1000, respectively.…”

Section: Methodsmentioning

confidence: 99%

Structural and Functional Consequences of Tyrosine Phosphorylation in the LRP1 Cytoplasmic Domain

Betts

Geer

Komives

2008

Journal of Biological Chemistry

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The low density lipoprotein receptor-related protein LRP1 is an endocytic receptor involved in the uptake of a diverse range of ligands including lipoproteins, proteases and protease-inhibitor complexes, matrix proteins, and growth factors (1). LRP1 is presented on the cell surface as a two-chain molecule, which consists of a 515-kDa ␣ chain and a 85-kDa ␤ chain that are noncovalently associated (2). The ␣ chain is made up of ligand-binding repeats organized in four domains, each resembling the extracellular domain of the low density lipoprotein receptor. The ligand-binding repeats have been shown to bind up to 30 different extracellular ligands, many of which are found in the proteinaceous plaques present in the brains of Alzheimer disease patients (3). The LRP1 cytoplasmic domain interacts with FE65, providing a direct link between LRP1 and the amyloid precursor protein (APP) 2 (4, 5).

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Section: Methodsmentioning

confidence: 99%

Structural and Functional Consequences of Tyrosine Phosphorylation in the LRP1 Cytoplasmic Domain

Betts

Geer

Komives

2008

Journal of Biological Chemistry

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“…Lucigenin (5 mg) was dissolved in HBSS to obtain a 10 -3 M solution. Further dilutions were in DMEM and the final concentration was 10 -5 M; 2) H 2 O 2 release was evaluated using peroxidase-catalysed CL oxidation of luminol [22,23]. Luminol (5 mg) was suspended in DMSO to obtain a 10 -2 M solution and further diluted in DMEM to a final concentration of 10 -4 M. Horseradish peroxidase (HRP) was added at a concentration of 0.2 U·10 -5 cells to optimize the reaction between luminol and H 2 O 2 .…”

Section: Oxygen Species Analysismentioning

confidence: 99%

Oxidant-antioxidant balance in alveolar macrophages from newborn rats

Delacourt¹,

D'Ortho²,

Macquin‐Mavier³

et al. 1996

Eur Respir J

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O Ox xi id da an nt t--a an nt ti io ox xi id da an nt t b ba al la an nc ce e i in n a al lv ve eo ol la ar r m ma ac cr ro op ph ha ag ge es s f fr ro om m n ne ew wb bo or rn n r ra at ts s ABSTRACT: An oxidant-antioxidant imbalance in neonatal alveolar macrophages (AMs) may contribute to the increased susceptibility to lung injury described in the neonatal period. We therefore evaluated oxygen radical production by rat AMs at various postnatal ages, and measured in parallel cellular antioxidant enzyme activities. AMs were obtained by bronchoalveolar lavage from rats aged <24 h, 21 days and 5 weeks, and results were compared to those obtained with adult rat AMs.Intracellular production of oxygen radical species, estimated fluorometrically using 2' ',5' '-dichlorofluorescein diacetate as the substrate, was significantly reduced in neonates as compared with adults, both in the presence and in the absence of cell stimulation with phorbol myristate acetate (PMA) or opsonized zymosan. A similar pattern was observed for the extracellular release of oxygen radical species, evaluated by lucigenin-enhanced chemiluminescence (CL) or peroxidase-catalysed CL oxidation of luminol: peak CL values measured after cell stimulation with PMA or opsonized zymosan remained significantly lower for AMs from newborn rats than for AMs from adults. By contrast, high values for antioxidant enzyme activities (superoxide dismutase and glutathione peroxidase) in AMs were demonstrated in newborns as compared to adults.We conclude that high antioxidant activity in rat AMs after birth may be at least partly responsible for the low production of oxygen metabolites observed during the same period.

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“…However, in the case of piodophenol, the phenoxy radical is thought to play an important role in the enhancing effect. 22,23 The decrease in the enhancing effect of p-iodophenylboronic acid or 4-biphenylboronic acid in the presence of saccharides was attributed to the interaction between the diol group of the saccharide and the boronic acid moiety of the enhancer. The interaction eliminated the possibility of generating such radicals.…”

Section: -Ch(oh)-ch(ch2oh)-mentioning

confidence: 99%

Enhancing Effect of Phenylboronic Acid Compounds and Their Interactions with the Diol Groups of Saccharides in a Capillary Electrophoresis-Chemiluminescence Detection System

et al. 2007

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Capillary electrophoresis(CE) equipped with chemiluminescence (CL) detection has attracted a great deal of attention as a high-performance separation and detection system. [1][2][3][4] We also analyzed many types of sample, including metal ions, metal compounds, amino acids, peptides, proteins, nucleic acids, saccharides, phenolic compounds, fluorescence compounds, and liposomes, using these systems. [5][6][7] We have determined the significant features of CE with CL detection systems through our previous research. 8,9 Naturally, while absorption and fluorescence detection in CE are carried out in an on-capillary manner, CL detection is brought about by endcapillary (postcolumn) detection. That is, CE with CL detection possesses a very interesting and useful "micro-environment area" for reaction/detection at the tip of the capillary outlet. Trace amounts of sample and reagents are delivered quantitatively and mixed reproducibly at the tip of the capillary outlet (micro-environment area), leading to useful and unique CL responses with high sensitivity.Saccharides are among the most important biological molecules in the living body. The compositional formulas of saccharides are simple, as they consist mainly of carbon, hydrogen, and oxygen. However, their molecular structures are very complicated. This complexity in the molecular structure is produced through the various directions of hydroxyl groups in the molecule. There have been many investigations of methodologies to derive information regarding the molecular structures of saccharides, i.e., molecular recognition of saccharides. 10,11 Boronic acids form cyclic esters with saccharides, particularly with those comprising cis-diol groups. [12][13][14] The interaction between boronic acids and saccharides has been used for molecular recognition of saccharides. To derive information regarding the interaction between boronic acids and saccharides, conventional procedures for molecular recognition require specific chemical reactions, such as polymerization and derivatization.In our previous study, 15 we found that the enhancing effect of p-iodophenylboronic acid on luminol-hydrogen peroxidehorseradish peroxidase CL reaction decreased in the presence of saccharide, and we proposed a system to take advantage of the micro-environment area of the CE-CL detection system for molecular recognition of saccharides. The decrease in CL could provide direct information regarding the interaction between piodophenylboronic acids and saccharides without requiring any specific procedures. The method for molecular recognition of saccharides has the following beneficial features in comparison with conventional methods, 16,17 including CD spectra, extraction, and π-A isotherm studies: 1) high sensitivity (a trace amount of a sample specimen is used), 2) rapid analysis, 3) ease of operation, and 4) inexpensive reagents and apparatus.In the present study, to extend our knowledge regarding the molecular recognition of saccharides using the CE with CL detection system, we examined the enh...

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[29] Enhanced chemiluminescent reactions catalyzed by horseradish peroxidase

Cited by 228 publications

References 30 publications

Structural and Functional Consequences of Tyrosine Phosphorylation in the LRP1 Cytoplasmic Domain

Structural and Functional Consequences of Tyrosine Phosphorylation in the LRP1 Cytoplasmic Domain

Oxidant-antioxidant balance in alveolar macrophages from newborn rats

Enhancing Effect of Phenylboronic Acid Compounds and Their Interactions with the Diol Groups of Saccharides in a Capillary Electrophoresis-Chemiluminescence Detection System

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