Facile determination of DNA-binding nuclear factor-κB by chemiluminescence detection

Tonooka, Keiko; Kabashima, Tsutomu; Yamasuji, Mutsumi; Kai, Masahiro

doi:10.1016/j.ab.2007.02.016

Cited by 10 publications

(7 citation statements)

References 19 publications

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“…Due to low solubility and instability of TCPO and other oxalate esters in an aqueous medium, the application of POCL in the detection of biological agents is very limited. A few publications have reported the detection of biological agents using POCL methods (21)(22)(23)(24). However, this reaction has to be performed in an organic solvent, or at least in an organic/aqueous mixed solvent (1:1 ratio), which is unacceptable for most biological reactions, given that they naturally need an aqueous environment.…”

Section: Introductionmentioning

confidence: 99%

Non‐enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

et al. 2008

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A new method for non-enzymatic aqueous peroxyoxalate chemiluminescence (POCL) biomolecular detection using imaging chip-based devices has been developed. A water-soluble amide of oxalic acid was synthesized and used in the investigation and characterization of POCL immunodetection in an aqueous environment. Six fluorescent dyes commonly used in biological detection were tested, and the intensity of light generated from the aqueous POCL reactions was characterized in the liquid phase. Direct detection sensitivity comparisons between a standard fluorescent method and this POCL method were performed in both liquid and solid phases. Results showed that detection sensitivity using the POCL method is comparable to that of the fluorescent method. POCL biomolecular detection on a nitrocellulose membrane was also investigated using a charge-coupled device (CCD) camera. Again, POCL detection sensitivity proved to be comparable to that using the fluorescent detection method. In an application of aqueous POCL biomolecular detection, Staphylococcus aureus enterotoxin B (SEB) and its antibody were used to demonstrate immuno- and affinity detection. For further applications, such as DNA and protein arrays, simultaneous detection of biomolecules labelled with different fluorescent labels was investigated, using a complementary metal oxide semiconductor (CMOS) colour imaging chip.

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Section: Introductionmentioning

confidence: 99%

Non‐enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

et al. 2008

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“…17 About 40 fmol (800 pg)/ 20 μL DNA with telomeric sequence (TTAGGG)10 could be quantified with a good reproducibility by the TMPG reaction at room temperature (24 -26˚C). The molecular sizes of the PCR products after the enzymatic reaction with telomerase were more than 45000 Da, and the molecular sizes of the substrate (= forward primer) and the reverse primer of the proposed method were less than 12000 Da.…”

Section: Telomerase Assay With CL Detectionmentioning

confidence: 99%

“…The TMPG reagent gives chemiluminescent signals selective for the guanine base in DNA, and is quantitatively determinable for the concentration of DNA in the reaction mixture. 17 The present method does not require any labeled probes and primers, and allows a sensitive analysis of the telomerase activity by a conventional chemiluminescence (CL) detector.…”

Section: Introductionmentioning

confidence: 99%

Facile Assay of Telomerase Activity Utilizing a DNA-detectable Chemiluminogenic Reagent

et al. 2008

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Telomerase shows increased activity in most human cancers and germ line cells, but not in normal human somatic cells. We describe a novel chemiluminescence method for the facile assay of telomerase activity in human cells. The telomerase substrate was incubated with the cell lysate containing various amounts of telomerase, and then the telomerase product was amplified by the polymerase-chained reaction (PCR). The PCR products were separated from the excess substrate, primer and deoxyribonucleotide triphosphates by a centrifugal filter, which distinguished different molecular sizes. The isolated products were reacted with a DNA-detectable chemiluminogenic reagent, 3,4,5-trimethoxyphenylglyoxal. The proposed assay method gave linearity for the telomerase activity in 100 to 10000 cells (r 2 = 0.997), and allowed the assay not only of lower activity, but also of higher activity of telomerase without the requirement of any special labeled-PCR primers in the assay system.

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“…Nuclear factor-kB (NF-kB), a sequence-specific DNA-binding protein, is a ubiquitous redox-sensitive transcription factor that responds to pro-inflammation caused by cytokines and oxidative stress. Tonooka et al 96 developed a facile method for the quantitative and sensitive determination of NF-kB by the TMPG-CL reaction. As shown in Fig.…”

Section: ·8·2 Protein Determinationmentioning

confidence: 99%

Chemiluminescence Platforms in Immunoassay and DNA Analyses

Fan

Cao

et al. 2009

ANAL. SCI.

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IntroductionCL is the production of electromagnetic radiation by a chemical reaction. The CL intensity is directly proportional to the concentration of a limiting reactant involved in the CL reaction. CL has been exploited for its use in a wide range of applications due to its high sensitivity, wide calibration range, and suitability for miniaturization. Protein and DNA analyses have recently attracted much attention. Many different types of techniques, such as electrochemical, optical, radiochemical, and piezoelectronic methods have been applied for protein and DNA analyses. To provide an overview of CL detection assays for DNAs and proteins, we have summarized their essential techniques. Monoplex and multiplex assays are described, and their current reports are intended to provide an overview of the CL techniques currently employed in numerous laboratories. Monoplex AssayA monoplex assay aims to measure the presence of a single analyte in any given sample, in which nanoparticle, DNA enzyme, horseradish peroxidase (HRP), alkaline phosphatase (AP) and acridinium ester are commonly used as labels. 2·1 Nanoparticle-based CL techniquesNanoparticles offer excellent prospects for DNA detection and immunoassay, owing to their many attractive properties. Compared to existing labels, nanoparticles are more stable and cheaper, and easier to be purified. In addition, they indicate faster binding kinetics with high sensitivity and a high reaction rate for many types of monoplex assays, ranging from immunoassays to DNA detection. 2·1·1 Gold nanoparticlesColloidal gold labels are ideal in biotechnological systems for several reasons. Firstly, they can be readily prepared in a wide range of sizes, from approximately 2 nm to above 100 nm. Secondly, the biochemical activity of the labeled biomolecules can be retained when colloidal gold is coupled to the biomolecules and, thirdly, colloidal gold labels can be easily visualized as dense structures within biological entities using transmission electron microscopy. As a biological tag, colloidal gold has been employed in several CL detection systems. 2·1·1·1 Gold nanoparticle-based CL immunoassay. The Au 3+ -luminol reaction, in which 10 -9 M Au 3+ ions can be sensitively detected, is a classic CL reaction. Thousands of Au atoms are contained within the gold nanoparticles; for example, 2.3 ¥ 10 5 Au atoms are theoretically contained in a 20-nm spherical gold particle and, consequently, picomolar detection limits can be attained by employing oxidative gold metal dissolution in acidic solutions. Our group 1,2 reported a sensitive CL immunoassay based on a colloidal gold label after oxidative gold metal dissolution (using 0.01 M HCl-0.5 M NaCl-0.5 mM Br2), using a simple and sensitive luminol-CL reaction (Fig. 1). Reviews Chemiluminescence Platforms in Immunoassay and DNA AnalysesAiping FAN,* Zhijuan CAO,* Huan LI,* Masaaki KAI,** and Jianzhong LU* † *School of Pharmacy, Fudan University, 138 Yixueyuan Road, Shanghai 200032, China ** Faculty of Pharmaceutical Sciences, Graduate Scho...

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Facile determination of DNA-binding nuclear factor-κB by chemiluminescence detection

Cited by 10 publications

References 19 publications

Non‐enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

Non‐enzymatic aqueous peroxyoxalate chemiluminescence immune detection using a CCD camera and a CMOS device

Facile Assay of Telomerase Activity Utilizing a DNA-detectable Chemiluminogenic Reagent

Chemiluminescence Platforms in Immunoassay and DNA Analyses

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