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Seed-Mediated Synthesis of Gold Nanorods: Role of …

A bacterial pellet (40 μg of total proteins) was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 12% linear gradient gels and then transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA). The membranes were first probed with an anti-MOMP monoclonal antibody (1:2000; Geneway Biotech, San Diego, CA), followed by a horseradish peroxidase-conjugated anti-mouse secondary antibody (1:2000; Amersham, UK). Immunoreactive proteins were detected using Luminol reagent (Santa Cruz Biotechnology, Santa Cruz, CA) and visualized by a Fuji Film image reader LAS-3000 machine.

The Advanced Laboratory for Iontronic, Electronic and Nanomaterials (ALIEN), led by Prof
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Chemiluminescence is another alternative energy source that serves as a FRET donor. For example, Luminol is a chemiluminescent reagent that is activated by oxidants such as H2O2. The reaction requires a catalyst to decompose H2O2 into H2O and O2. The hemin/G-quadruplex horseradish peroxidase (HRP) mimicking catalytic nucleic acids (DNAzymes) were discovered to generate chemiluminescence through catalyzing the oxidation of luminol by H2O2 [, ]. Freeman included aptamer domains into the DNAzyme subunits []. One of the aptamer subunits was conjugated to a QD. In the presence of aptamer targets, ATP or Hg2+ in this case, the DNAzyme subunits self assembled into active hemin/G-quadruplex DNAzyme structures and promoted the chemiluminescence resonance energy transfer (CRET) by catalyzing luminol emission. As shown in Figure A, nucleic subunits included domain I and II of the HRP mimicking DNAzyme, as well as domain V and VI of an anti-ATP aptamer. In the absence of ATP, the two subunits are not able to form a stable complex. However, in the presence of ATP, the aptamer domains binds to ATP and the resulting complex leads to the formation of a hemin/G-quadruplex that catalyzes the chemiluminescent reaction and gives rise to CRET. In contrast to FRET, the emission intensities of donors and acceptors increased or decreased concurrently because the amount of energy transferred to the QD was proportional to the chemiluminescent energy available (Fig. B). QD-CRET sensors were also configured to detect specific DNA sequences (Fig. C). A DNA hairpin structure consisting of a few functional domains were conjugated to QDs. The DNAzyme forming domain was blocked in the presence of the hairpin loop. The sequence recognition domain resided in the loop. As the target DNA hybridized to the recognition sequence and opened the hairpin, the DNAzyme forming domain was freed, leading to the self assembly of a hemi/G-quadruplex DNAzyme. DNA hairpins with three different target recognition sequences were conjugated to QD490, QD560 and QD620 respectively to form three QD-CRET DNA probes. Upon hybridization to their respective targets, Hemin and H2O2 were added to induce CRET. The presence of targets was indicated by emission of specific QDs probes through CRET. With the proposed QD-CRET sensor, the authors successfully resolved three targets in a multiplexed format (Fig. D).

Regional Centre of Advanced Technologies and Materials

Chemiluminescence of Luminol in the Presence of Iron(II) and Oxygen: Oxidation Mechanism and Implications for Its Analytical Use Andrew L
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In addition to environmental conditions, QD emission intensities are strongly influenced by proximal molecules or nanoparticles that QDs interact with. Under many circumstances, the photoluminescence of QDs is drastically quenched through numerous mechanisms. This seemingly undesirable phenomenon can serve as an advantageous feature. For example, quenching mechanism can be designed to act like a molecular switch for fluorescent signals, which would make QDs an ideal homogeneous sensing platform for studying molecular interactions and detecting specific targets.

Researche institute in Czech Republic focuses on nanotechnology

Chemiluminescence of luminol in the presence of Iron(II) and oxygen: oxidation mechanism and implications for Its analytical use
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Recent Advances in the Synthesis and Application of Layered Double Hydroxide (LDH) Nanosheets
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