No (118)
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Separation of tryptic digest of cytochrome C within a long nanoelectrospray tip
Týčová, Anna; Foret, František
2013 - English
Capillary electrophoresis coupled to mass spectrometry (CE-MS) posses powerful tool for separation and detection of wide range of ionic species. To avoid complicated constructions of interfaces we have conducted the separation within a long and thin nanoelectrospray tip. We have investigated main properties of such an experimental design on a complex sample of tryptic digest of cytochrome c.
Keywords:
cytochrome c; thin nanoelectrospray tip; electrophoretic separation
Available on request at various institutes of the ASCR
Separation of tryptic digest of cytochrome C within a long nanoelectrospray tip
Capillary electrophoresis coupled to mass spectrometry (CE-MS) posses powerful tool for separation and detection of wide range of ionic species. To avoid complicated constructions of interfaces we ...
New isoelectric focusing power supply based on features of voltage multiplier
Duša, Filip; Šlais, Karel
2013 - English
Present electrophoretic separation methods rely on sophisticated high-voltage power supplies capable of programing a voltage / current time course during an analysis. In this paper we suggest design of a simple highvoltage power supply for isoelectric focusing composed from affordable and commonly available electrical parts. It is based on features of the voltage multiplier invented by Cockcroft and Walton. Electrical characteristics of the power supply enabled power load controlled isoelectric focusing analysis thus eliminating need for programing voltage time course and reducing analysis total time.
Keywords:
isoelectric focusing; IEF device; electrodes
Available on request at various institutes of the ASCR
New isoelectric focusing power supply based on features of voltage multiplier
Present electrophoretic separation methods rely on sophisticated high-voltage power supplies capable of programing a voltage / current time course during an analysis. In this paper we suggest ...
Thin metal films for detection and preconcentration
Podešva, Pavel; Foret, František
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:
thin metal film; chemoresistor; Au nanoparticles
Available on request at various institutes of the ASCR
Thin metal films for detection and preconcentration
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 ...
Utilization of micropreparative fast focusing by a new wide pH range electrolyte system based on bidirectional isotachophoresis
Vykydalová, Marie; Duša, Filip; Horká, Marie; Šlais, Karel
2013 - English
We suggest the possibility of practical utilization of a new electrolyte system for fast preparative focusing in wide pH range based on bidirectional isotachophoresis. The focusing occurs on nonwoven fabric strip positioned in an open horizontal V-shaped trough. It is based on bidirectional ITP with multiple counter ions and spacers created from commercially available simple buffers. Milk spiked with ampicillin was used as a sample and high performance liquid chromatography was as a second dimension for analysis of fast preparative focusing fractions. Speed, easy fraction handling, and possibility of pre-concentration of analytes from a raw sample are the benefits of this technique.
Keywords:
bidirectional isotachophoresis; electrolyte system; preparative focusing
Available on request at various institutes of the ASCR
Utilization of micropreparative fast focusing by a new wide pH range electrolyte system based on bidirectional isotachophoresis
We suggest the possibility of practical utilization of a new electrolyte system for fast preparative focusing in wide pH range based on bidirectional isotachophoresis. The focusing occurs on ...
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 ...
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