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Search results for: Cy3

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#32764941   2020/07/16 To Up

Efficient miRNA Inhibitor with GO-PEI Nanosheets for Osteosarcoma Suppression by Targeting PTEN.

Gene therapy is considered a novel way to treat osteosarcoma, and microRNAs are potential therapeutic targets for osteosarcoma. miR-214 has been found to promote osteosarcoma aggression and metastasis. Graphene oxide (GO) is widely used for gene delivery for the distinct physiochemical properties and minimal cytotoxicity.
Lingling Ou, Haiyingjie Lin, Yuwei Song, Guoqiang Tan, Xiujuan Gui, Jinyuan Li, Xiaoting Chen, Zhendong Deng, Shaoqiang Lin

2752 related Products with: Efficient miRNA Inhibitor with GO-PEI Nanosheets for Osteosarcoma Suppression by Targeting PTEN.

50ul5mg100 assays1 LITRE100 µl5 mg100 μg400Tests5 mg5mg100

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#32661869   2020/07/13 To Up

Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells.

Sperm-mediated gene transfer (SMGT) has a potential use for zebrafish transgenesis. However, transfection into fish sperm cells still needs to be improved. The objective was to demonstrate the feasibility of tip type electroporation in zebrafish sperm, showing a protocol that provide high transfection efficiency, with minimal side-effects. Sperm was transfected with a Cy3-labelled DNA using tip type electroporation with voltages ranging from 500 to 1500 V. Sperm kinetics parameters were assessed using Computer Assisted Semen Analysis (CASA) and cell integrity, reactive oxygen species (ROS), mitochondrial functionality and transfection rate were evaluated by flow cytometry. The transfection rates were positively affected by tip type electroporation, reaching 64.9% ± 3.6 in the lowest voltage used (500 V) and 86.6% ± 1.9 in the highest (1500 V). The percentage of overall motile sperm in the electrotransfected samples was found to decrease with increasing field strength (P < 0.05). Increase in the sperm damaged plasma membrane was observed with increasing field strength (P < 0.05). ROS and sperm mitochondrial functionality did not present a negative response after the electroporation (P > 0.05). Overall results indicate that tip type electroporation enhances the internalization of exogenous DNA into zebrafish sperm cells with minimal harmful effects to sperm cells.
Larissa O Daneluz, Izani B Acosta, Leandro S Nunes, Eduardo B Blodorn, William B Domingues, Amanda W S Martins, Eduardo N Dellagostin, Gabriela T Rassier, Carine D Corcini, Charles N Fróes, Eliza R Komninou, Antônio S Varela, Vinicius F Campos

1409 related Products with: Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells.

1 mg1.00 flask10 rxns1x10e7 cells1.00 flask96 tests1x10e7 cells-96 tests1x10e7 cells100 µg

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#32648544   2020/07/10 To Up

Real-time in vivo imaging of extracellular ATP in the brain with a hybrid-type fluorescent sensor.

Adenosine 5' triphosphate (ATP) is a ubiquitous extracellular signaling messenger. Here, we describe a method for in-vivo imaging of extracellular ATP with high spatiotemporal resolution. We prepared a comprehensive set of cysteine-substitution mutants of ATP-binding protein, FoF-ATP synthase ε subunit, labeled with small-molecule fluorophores at the introduced cysteine residue. Screening revealed that the Cy3-labeled glutamine-105 mutant (Q105C-Cy3; designated ATPOS) shows a large fluorescence change in the presence of ATP, with submicromolar affinity, pH-independence, and high selectivity for ATP over ATP metabolites and other nucleotides. To enable in-vivo validation, we introduced BoNT/C-Hc for binding to neuronal plasma membrane and Alexa Fluor 488 for ratiometric measurement. The resulting ATPOS complex binds to neurons in cerebral cortex of living mice, and clearly visualized a concentrically propagating wave of extracellular ATP release in response to electrical stimulation. ATPOS should be useful to probe the extracellular ATP dynamics of diverse biological processes in vivo.
Nami Kitajima, Kenji Takikawa, Hiroshi Sekiya, Kaname Satoh, Daisuke Asanuma, Hirokazu Sakamoto, Shodai Takahashi, Kenjiro Hanaoka, Yasuteru Urano, Shigeyuki Namiki, Masamitsu Iino, Kenzo Hirose

2303 related Products with: Real-time in vivo imaging of extracellular ATP in the brain with a hybrid-type fluorescent sensor.

96 tests2596 tests2596 tests2596 tests2596T

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#32647943   2020/07/09 To Up

"Plug and Play" logic gate construction based on chemically triggered fluorescence switching of gold nanoparticles conjugated with Cy3-tagged aptamer.

Gold nanoparticles (AuNPs) conjugated with Cy3-tagged aptamer which can specifically recognize chloramphenicol (CAP) (referred to as AuNPs-AptCAP) are described. CAP can trigger the configuration change of CAP binding aptamer, and thus switching the fluorescence of AuNPs-AptCAP through changing the efficiency of the fluorescence resonance energy transfer (FRET) system with Cy3 as donors and AuNPs as recipients. AuNPs-AptCAP exhibits a linear range of CAP concentrations from 26.0 to 277 μg L with a limit of detection of 8.1 μg L when Cy3 was excited at 530 nm and emission was measured at 570 nm. More importantly, AuNPs-AptCAP can be utilized as signal transducers for the build-up of a series of logic gates including YES, PASS 0, INH, NOT, PASS 1, and NAND. Utilizing the principle of a metal ion-mediated fluorescence switch together with a strong metal ion chelator, the fluorescence of AuNPs-AptCAP could be modulated by adding metal ions and EDTA sequentially. Therefore, a "Plug and Play" logic system based on AuNPs-AptCAP has been realized by simply adding other components to create new logic functions. This work highlights the advantages of simple synthesis and facile fluorescence switching properties, which will provide useful knowledge for the establishment of molecular logic systems. Graphical abstract.
Yali Zhang, Cheuk-Wing Li, Lefei Zhou, Zhanpeng Chen, Changqing Yi

2491 related Products with: "Plug and Play" logic gate construction based on chemically triggered fluorescence switching of gold nanoparticles conjugated with Cy3-tagged aptamer.

100ug100ug Lyophilized100ug Lyophilized100ug100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug100ug Lyophilized

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#32614024   2020/07/02 To Up

Supramolecular assembly of DNA-constructed vesicles.

The self-assembly of DNA hybrids possessing tetraphenylethylene sticky ends at both sides into vesicular architectures in aqueous medium is demonstrated. Cryo-electron microscopy reveals the formation of different types of morphologies from the amphiphilic DNA-hybrids. Depending on the conditions, either an extended (sheet-like) or a compact (columnar) alignment of the DNA hybrids is observed. The different modes of DNA arrangement lead to the formation of vesicles appearing either as prolate ellipsoids (type I) or as spheres (type II). The type of packing has a significant effect on the accessibility of the DNA, as evidenced by intercalation and light-harvesting experiments. Only the vesicles exhibiting the sheet-like DNA alignment are accessible for intercalation by ethidium bromide or for the integration of chromophore-labelled DNA via a strand exchange process. The dynamic nature of type I vesicles enables their elaboration into artificial light-harvesting complexes by DNA-guided introduction of Cy3-acceptor chromophores. DNA-constructed vesicles of the kind shown here represent versatile intermediates that are amenable to further modification for tailored nanotechnology applications.
Simon Rothenbühler, Ioan Iacovache, Simon M Langenegger, Benoît Zuber, Robert Häner

2328 related Products with: Supramolecular assembly of DNA-constructed vesicles.

100ul100ul100ug100ug 5 G

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#32594743   2020/07/13 To Up

Unifying Mechanism for Thiol-Induced Photoswitching and Photostability of Cyanine Dyes.

Cyanines (Cy3, Cy5, Cy3B) are the most utilized dyes for single-molecule fluorescence and localization-based super-resolution imaging. These modalities exploit cyanines' versatile photochemical behavior with thiols. A mechanism reconciling seemingly divergent results and enabling control over cyanine photoreactivity is however missing. Utilizing single-molecule fluorescence on Cy5 and Cy5B, transient-absorption spectroscopy, and DFT modeling on a range of cyanine dyes, herein we show that photoinduced electron transfer (PT) from a thiolate to Cy in their triplet excited state and then triplet-to-singlet intersystem crossing in the nascent geminate radical pair are crucial steps. Next, a bifurcation occurs, yielding either back electron transfer and regeneration of ground state Cy, required for photostabilization, or Cy-thiol adduct formation, necessary for super-resolution microscopy. Cy regeneration via photoinduced thiol elimination is favored by adduct absorption spectra broadening. Elimination is also shown to occur through an acid-catalyzed reaction. Overall, our work provides a roadmap for designing fluorophores, photoswitching agents, and triplet excited state quenchers for single-molecule and super-resolution imaging.
Yasser Gidi, Liam Payne, Viktorija Glembockyte, Megan S Michie, Martin J Schnermann, Gonzalo Cosa

1405 related Products with: Unifying Mechanism for Thiol-Induced Photoswitching and Photostability of Cyanine Dyes.

5mg5mg100Tests5mg200ul 1 G5mg 2x5L5mg25 mg50 ul

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#32533943   2020/06/02 To Up

Membrane Interactions of Cy3 and Cy5 Fluorophores and Their Effects on Membrane-Protein Dynamics.

Organic fluorophores, such as Cy3 and Cy5, have been widely used as chemical labels to probe the structure and dynamics of membrane proteins. Although a number of previous studies have reported on the possibility of some of the water-soluble fluorophores to interact with lipid bilayers, detailed fluorophore-lipid interactions and, more importantly, the potential effect of such interactions on the natural dynamics of the labeled membrane proteins have not been well studied. We have performed a large set of all-atom molecular dynamics simulations employing the highly mobile membrane mimetic model to describe spontaneous partitioning of the fluorophores into lipid bilayers with different lipid compositions. Spontaneous membrane partitioning of Cy3 and Cy5 fluorophores captured in these simulations proceeds in two steps. Electrostatic interaction between the fluorophores and the lipid headgroups facilitates the initial, fast membrane association of the fluorophores, followed by slow insertion of hydrophobic moieties into the lipid bilayer core. After the conversion of the resulting membrane-bound systems to full-membrane representations, biased-exchange umbrella sampling simulations are performed for free energy calculations, revealing a higher energy barrier for partitioning into negatively charged (phosphatidylserine or phosphatidylcholine) membranes than purely zwitterionic (phosphatidylcholine or phosphatidylethanolamine) ones. Furthermore, the potential effect of fluorophore-lipid interactions on membrane proteins has been examined by covalently linking Cy5 to single- and multipass transmembrane helical proteins. Equilibrium simulations show strong position-dependent effects of Cy5-tagging on the structure and natural dynamics of membrane proteins. Interactions between the tagged protein and Cy5 were also observed. Our results suggest that fluorophore-lipid interactions can affect the structure and dynamics of membrane proteins to various extents, especially in systems with higher structural flexibility.
Kin Lam, Emad Tajkhorshid

1298 related Products with: Membrane Interactions of Cy3 and Cy5 Fluorophores and Their Effects on Membrane-Protein Dynamics.

100ul0.1 mg0.1 mg250 100ul1000 TESTS/0.65ml100ul100μg100 100μg50 assays

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#32520054   // To Up

Impact of non-nucleotidic bulges on recognition of mixed-sequence dsDNA by pyrene-functionalized Invader probes.

Invader probes, i.e., DNA duplexes modified with +1 interstrand zippers of intercalator-functionalized nucleotides like 2'-O-(pyren-1-yl)methyl-RNA monomers, are energetically activated for sequence-unrestricted recognition of double-stranded DNA (dsDNA) as they are engineered to violate the neighbor exclusion principle, while displaying high affinity towards complementary DNA sequences. The impact on Invader-mediated dsDNA-recognition upon additional modification with different non-nucleotidic bulges is studied herein, based on the hypothesis that bulge-containing Invader probes will display additionally disrupted base-stacking, more extensive denaturation, and improved dsDNA-recognition efficiency. Indeed, Invader probes featuring a single central large bulge - e.g., a nonyl (C9) monomer - display improved recognition of model DNA hairpin targets vis-à-vis conventional Invader probes (C50 values ∼1.5 μM vs. ∼3.9 μM). In contrast, probes with two opposing central bulges display less favorable binding characteristics. Remarkably, C9-modified Invader probes display perfect discrimination between fully complementary dsDNA and dsDNA differing in only one of eighteen base-pairs, underscoring the high binding specificity of double-stranded probes. Cy3-labeled bulge-containing Invader probes are demonstrated to signal the presence of gender-specific DNA sequences in fluorescent in situ hybridization assays (FISH) performed under non-denaturing conditions, highlighting one potential application of dsDNA-targeting Invader probes.
Dale C Guenther, Raymond G Emehiser, Allison Inskeep, Saswata Karmakar, Patrick J Hrdlicka

2255 related Products with: Impact of non-nucleotidic bulges on recognition of mixed-sequence dsDNA by pyrene-functionalized Invader probes.

5 G 2 ml Ready-to-use 100 ug1 mg10reactions 100 µg25 mg0.2 mg100 ug/vial10ml1 mg 6 ml Ready-to-use

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#32510543   // To Up

Octahedral copper(ii)-diimine complexes of triethylenetetramine: effect of stereochemical fluxionality and ligand hydrophobicity on Cu/Cu redox, DNA binding and cleavage, cytotoxicity and apoptosis-inducing ability.

Octahedral copper(ii) complexes of the type [Cu(trien)(diimine)](ClO4)2 (1-4), where trien is triethylenetetramine and diimine is 2,2'-bipyridine (1), 1,10-phenanthroline (2), 5,6-dimethyl-1,10-phenanthroline (3), and 3,4,7,8-tetramethyl-1,10-phenanthroline (4), have been isolated. Single crystal X-ray structures of 1 and 2 reveal that the coordination geometry around Cu(ii) is tetragonally distorted octahedral. The stereochemical fluxionality of the complexes illustrates the observed trend in CuII/CuI redox potentials and DNA binding affinity (Kb: 1, 0.030 ± 0.002 < 2, 0.66 ± 0.01 < 3, 1.63 ± 0.10 < 4, 2.27± 0.20 × 105 M-1), determined using absorption spectral titration. All complexes effect oxidative DNA cleavage more efficiently than hydrolytic DNA cleavage. The bpy complex 1 with stereochemical fluxionality lower than its phen analogue 2 shows a higher cytotoxicity against both A549 lung (IC50, 3.3 μM) and MCF-7 human breast (IC50, 3.9 μM) cancer cells, and induces the generation of the highest amount of ROS in A549 cells. Complex 3 with a higher stereochemical fluxionality and higher ligand hydrophobicity exhibits a higher DNA binding and cleavage ability and higher cytotoxicity (IC50, 2.1 μM) towards MCF-7 cells. Complex 4 with a still higher stereochemical fluxionality displays the highest DNA binding and cleavage ability but a lower cytotoxicity towards both A549 and MCF-7 cell lines due to its tendency to form a five-coordinated complex with the uncoordinated amine group. Annexin V.Cy3 staining and immunoblot analysis demonstrate the mechanism of cell death caused by 1 and 2. The finding of the up-regulation of the pro-apoptotic Bax protein and down-regulation of PARP protein in western blot analysis confirms the induction of apoptosis by these complexes.
Mitu Sharma, Mani Ganeshpandian, Munmi Majumder, Ajaykamal Tamilarasan, Mukesh Sharma, Rupak Mukhopadhyay, Nashreen S Islam, Mallayan Palaniandavar

2682 related Products with: Octahedral copper(ii)-diimine complexes of triethylenetetramine: effect of stereochemical fluxionality and ligand hydrophobicity on Cu/Cu redox, DNA binding and cleavage, cytotoxicity and apoptosis-inducing ability.

1000 TESTS/0.65ml100ul500 MG100ug25 mg10 mg100ug50 ug 1 ml50 mg100ug25 mg

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