Multi-step synthesis of caspase-3 sensor based on Förster resonance energy transfer
Lišková, Marcela; Klepárník, Karel; Pazdera, P.; Foret, František
2013 - English
Programmed cell death or apoptosis is regulated process of cell suicide. The central role in apoptosis play cysteine proteases called caspases. Caspases recognize tetra-peptide sequences Asp-Glu-Val-Asp (DEVD) on their substrates and hydrolyze peptide bonds after aspartic acid residues. Various techniques for the determination of caspase-3 are commercially available e.g. Enzyme Linked Immuno-Sorbent Assay (ELISA), Western blotting or flow cytometric analysis. The products of the cleavage can be detected by spectrophotometry, fluorimetry, chemiluminescence (CL) or ELISA. In this work, we suggested fluorescent sensor based on Förster Resonance Energy Transfer (FRET) to determine caspase-3 in cell nucleus or cytoplasm by apoptosis for very fast analysis by fluorescence microscopy without sample destroying.
Keywords:
caspase-3; Förster resonance energy transfer; apoptotic cells
Available on request at various institutes of the ASCR
Multi-step synthesis of caspase-3 sensor based on Förster resonance energy transfer
Programmed cell death or apoptosis is regulated process of cell suicide. The central role in apoptosis play cysteine proteases called caspases. Caspases recognize tetra-peptide sequences ...
Capillary isotachophoresis of anions with electrospray-ionization mass-spectrometric detection
Malá, Zdeňka; Gebauer, Petr; Boček, Petr
2013 - English
We present ITP-ESI-MS as a powerful tool for selective and sensitive analyses of anionic analytes. Our extended concept of ITP includes besides regular and freeacid ITP also self-maintained ITP in moving boundary systems. Easy theory describes the system properties including their stacking-window diagrams. Simple ESIcompatible electrolyte systems allow flexible tuning with very satisfactory results as demonstrated on the example of analysis of pharmaceuticals in waters.
Keywords:
capillary isotachophoresis; electrospray ionization; mass-spectrometric detection
Available on request at various institutes of the ASCR
Capillary isotachophoresis of anions with electrospray-ionization mass-spectrometric detection
We present ITP-ESI-MS as a powerful tool for selective and sensitive analyses of anionic analytes. Our extended concept of ITP includes besides regular and freeacid ITP also self-maintained ITP in ...
Polydopamine coated capillaries for CE separations
Partyka, Jan; Foret, František; Vu, T. T.; Sudor, J.
2013 - English
We have tested CE separation of selected samples in capillaries with polydopamine modified surface. The capillaries were modified by polydopamine or polydopamine with an additive. The polydopamine coating with additive represents a simple and effective procedure for capillary alteration by another modificator such as hydroxyethyl cellulose, hydroxypropyl cellulose etc. In this work, we represent separation data from PrinCE system with UV detection for peptides, proteins and oligosaccharides labeled by 2-aminobenzoic acid.
Keywords:
CE separations; polydopamine; hydroxyethyl cellulose
Available on request at various institutes of the ASCR
Polydopamine coated capillaries for CE separations
We have tested CE separation of selected samples in capillaries with polydopamine modified surface. The capillaries were modified by polydopamine or polydopamine with an additive. The polydopamine ...
Integrated microfluidic device for droplet manipulation
Basova, E.; Drs, J.; Zemánek, J.; Hurák, Z.; Foret, František
2013 - English
Droplets based microfluidic systems have a big potential for the miniaturization of processes for bioanalysis. In the form of droplets, reagents are used in discrete volume, enabling high-throughput chemical reactions as well as single-cell encapsulation. Microreactors of this type can be manipulated and applied in bio-testing. In this work we present a platform for droplet generation and manipulation by using dielectrophoresis force. This platform is an integrated microfluidic device with a dielectrophoresis (DEP) chip. The microfluidic device generates microdroplets such as water in oil emulsion.
Keywords:
microfluidic device; miniaturization; bioanalysis; droplets
Available on request at various institutes of the ASCR
Integrated microfluidic device for droplet manipulation
Droplets based microfluidic systems have a big potential for the miniaturization of processes for bioanalysis. In the form of droplets, reagents are used in discrete volume, enabling high-throughput ...
Porous layer open tubular columns with immobilized trypsin for protein digestion
Knob, R.; Křenková, Jana; Petr, J.; Foret, František
2013 - English
We have developed a monolithic porous layer open tubular (PLOT) column with immobilized trypsin for protein digestion. The PLOT column was prepared in a 10 (mikro)m ID fused silica capillary. Trypsin was immobilized on the monolithic surface and the developed enzyme reactor was used for protein digestion followed by on-line ESI/MS analysis.
Keywords:
monolith; enzymes; digestion
Available on request at various institutes of the ASCR
Porous layer open tubular columns with immobilized trypsin for protein digestion
We have developed a monolithic porous layer open tubular (PLOT) column with immobilized trypsin for protein digestion. The PLOT column was prepared in a 10 (mikro)m ID fused silica capillary. ...
High pressure modification of the simple automated liquid chromatographic system for splitless nano column gradient separations
Šesták, Jozef; Kahle, Vladislav
2013 - English
Our simple liquid chromatographic system for splitless gradient nanocolumn separations at backpressure up to 50 bars was modified to run analysis under the pressure up to 300 bars. Programmable syringe pump was equipped with a 250-L high-pressure syringe and mobile phase gradients were created and stored in relatively long capillary. Available linear gradients of volumes from 5 to 50 L traced by uracil are presented. Sample introduction was performed as separate step and real large volume of sample (tens of microliters) could be injected and preconcentrated on-column. Relative standard deviation of retention times and peak areas were received (RSD < 0.3 % and RSD < 5 % respectively).
Keywords:
liquid chromatography; nano column gradient separations; gradient separation
Available on request at various institutes of the ASCR
High pressure modification of the simple automated liquid chromatographic system for splitless nano column gradient separations
Our simple liquid chromatographic system for splitless gradient nanocolumn separations at backpressure up to 50 bars was modified to run analysis under the pressure up to 300 bars. Programmable ...
The first experiencies with new exposure system for the whole body inhalation experiments with small animals
Večeřa, Zbyněk; Mikuška, Pavel; Moravec, Pavel; Smolík, Jiří
2013 - English
The first experiences with an inhalation chamber constructed for long-term exposure studies of small animals to nanoparticles under strictly controlled experimental conditions. The inhalation chamber was build from a glass and stainless steel and allows possible long-term inhalation procedures with up to four discrete groups of mice or of rats population under controlled illumination, temperature, relative humidity and pressure. Doses up to 1 x 10(6) particles/cm(3) and size (10-60 nm) of nano-sized particles were synthesized via aerosol route in a hot wall tube flow reactor using thermal decomposition of metal organic precursors. The function of exposure system for whole body inhalation experiments was studied under real conditions in period from May to September 2012. No animals were used within this trial experiment.
Keywords:
inhalation chamber; nanoparticles; small animals
Available at various institutes of the ASCR
The first experiencies with new exposure system for the whole body inhalation experiments with small animals
The first experiences with an inhalation chamber constructed for long-term exposure studies of small animals to nanoparticles under strictly controlled experimental conditions. The inhalation chamber ...
New sensor for DNA mutation detection
Datinská, Vladimíra; Klepárník, Karel; Minárik, M.; Foret, František
2013 - English
We present a design and synthesis of a new sensor intended for the genomic analysis in molecular cancer research. This sensor is based on the Förster resonance energy transfer (FRET) and can be used for the detection of complementary oligonucletide chains based on probe hybridization. Quantum dots (QDs) with their unique optical properties serve as suitable donors of energy in FRET.
Keywords:
quantum dots; Förster resonance energy transfer; CE-LIF
Available on request at various institutes of the ASCR
New sensor for DNA mutation detection
We present a design and synthesis of a new sensor intended for the genomic analysis in molecular cancer research. This sensor is based on the Förster resonance energy transfer (FRET) and can be ...
IEF and HPLC-based methods for efficient bacterial characterization
Kubesová, Anna; Šalplachta, Jiří; Moravcová, Dana; Horká, Marie
2013 - English
Pectobacterium and Dickeya species are responsible for blackleg and tuber soft rot diseases on crop and ornamental plants. Recently, a more virulent and aggressive Dickeya type of E. chrysanthemi was described and it becomes serious problem for potato production in Europe. In this study, we suggest procedures using two commonly available laboratory techniques, gel isoelectric focusing and liquid chromatography, for differentiation and characterization of Pectobacterium and Dickeya species. We have confirmed that fingerprinting approach can be used for bacterial differentiation, which can be further used in rapid diagnosis of plant disease.
Keywords:
Pectobacterium; Dickeya species; gel isoelectric focusing; liquid chromatography
Available on request at various institutes of the ASCR
IEF and HPLC-based methods for efficient bacterial characterization
Pectobacterium and Dickeya species are responsible for blackleg and tuber soft rot diseases on crop and ornamental plants. Recently, a more virulent and aggressive Dickeya type of E. chrysanthemi ...
Electrophoretic mobility measurements of a multivalent red dye SPADNS
Pantůčková, Pavla; Kubáň, Pavel; Boček, Petr
2013 - English
Measurements of the electrophoretic mobility of an anionic dye 4,5-dihydroxy-3-(p-sulfophenylazo)-2,7- naphthalene disulfonic acid trisodium salt (SPADNS), which is a commonly used anionic dye, are described in this contribution. Electrophoretic mobilities of SPADNS were measured in various buffer solutions covering the pH range of 2.4–9.1. Effective electrophoretic mobilities of SPADNS measured in these buffers varied between 50.5 x 10–9 m2/Vs and 60.5 x 10–9 m2/Vs with an average effective mobility of 55.0 x 10–9 m2/Vs.
Keywords:
electrophoretic mobility; SPADNS; electrophoretic separations
Available on request at various institutes of the ASCR
Electrophoretic mobility measurements of a multivalent red dye SPADNS
Measurements of the electrophoretic mobility of an anionic dye 4,5-dihydroxy-3-(p-sulfophenylazo)-2,7- naphthalene disulfonic acid trisodium salt (SPADNS), which is a commonly used anionic dye, are ...
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