Minimal Information About Sample Preparation for Phosphoproteomics

This guideline describes parameters and conditions involved in phosphopeptide sample preparation. It covers from the description and preparation of the cells and tissues to the fractionation and specific enrichment of phosphopeptides for MS analysis. The guideline is prepared in order to easily cope with many of the experimental designs used in phosphoproteomic studies. 
 
The document is subdivided as follows:
1. General features
2. Sample processing
3. Protein Purification/Fractionation
4. Peptide Purification/Fractionation
5. Phosphopeptide enrichment
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Sample preparation for point of care molecular diagnostics of STIs

This paper was presented at the 4th Micro and Nano Flows Conference (MNF2014), which was held at University College, London, UK. The conference was organised by Brunel University and supported by the Italian Union of Thermofluiddynamics, IPEM, the Process Intensification Network, the Institution of Mechanical Engineers, the Heat Transfer Society, HEXAG - the Heat Exchange Action Group, and the Energy Institute, ASME Press, LCN London Centre for Nanotechnology, UCL University College London, UCL Engineering, the International NanoScience Community, www.nanopaprika.eu.Brunel DoCLab is part of the esti2 consortium developing electronic self-testing instruments for sexually transmitted infections using nucleic acid amplification testing (NAAT). A proprietary sample collection device has been designed to integrate directly with a microfluidic cartridge. Cell lysis was conducted using a chemical method and nucleic acid purification was done on an activated cellulose membrane. The microfluidic device incorporates passive mixing of the lysis-binding buffers and sample. Preliminary results have shown extraction efficiencies for this new membrane of 69% and 57% compared to the commercial Qiagen extraction method of 85% and 59.4% for 0.1ng/μL and 100ng/μL salmon sperm DNA spiked in phosphate buffered solution. Preliminary extraction experiments in the passive mixer cartridges with lysis and nucleic acid purification showed extraction efficiency around 80% of the commercial Qiagen kit. Isothermal amplification was conducted using thermophillic helicase dependant amplification. A low cost benchtop real-time isothermal amplification platform has been developed capable of running six amplifications simultaneously. Work to integrate sample collection, nucleic acid extraction and isothermal amplification is currently underway

Sample Preparation for Determination of Biological Thiols by Liquid Chromatography and Electromigration Techniques

Wydrukowano z dostarczonych Wydawnictwu UŁ gotowych materiałówMajority of the bioanalytical or environmental methods do not use just one chromatografie or electrophoretic step, but rather involve several sample pretreatment steps which simplfy the matrix, and often preconcentrate and chemically modify the analytes. This work surveys typical procedures for sample preparation for most commonly analyzed biofluids with particular emphasis placed on chemical derivatization of sulfur amino acids for their determination by liquid phase separation techniques. Recent author's laboratory contribution to the development of sample preparation procedures is merked.Zadanie pt. Digitalizacja i udostępnienie w Cyfrowym Repozytorium Uniwersytetu Łódzkiego kolekcji czasopism naukowych wydawanych przez Uniwersytet Łódzki nr 885/P-DUN/2014 zostało dofinansowane ze środków MNiSW w ramach działalności upowszechniającej naukę

Sample preparation of metal alloys by electric discharge machining

Electric discharge machining was investigated as a noncontaminating method of comminuting alloys for subsequent chemical analysis. Particulate dispersions in water were produced from bulk alloys at a rate of about 5 mg/min by using a commercially available machining instrument. The utility of this approach was demonstrated by results obtained when acidified dispersions were substituted for true acid solutions in an established spectrochemical method. The analysis results were not significantly different for the two sample forms. Particle size measurements and preliminary results from other spectrochemical methods which require direct aspiration of liquid into flame or plasma sources are reported

Datasets: Sensitivity and protein digestion course of proteomic Filter Aided Sample Preparation

Sensitivity of FASP was tested using SDS lysates from HeLa cells and mouse brain. Peptides were analyzed using a QExactive HF-X instrument. Whole cell lysates of Hela cells were processed with FASP using single or double, consecutive or successive, digestion with LysC or trypsin. The generated peptides were analyzed using a LTQ-Orbitrap mass spectrometer. These datasets accompany "Filter Aided Sample Preparation - A Tutorial" (Wisniewski, 2019). (C) 2019 The Authors. Published by Elsevier Inc

Uncertainties of size measurements in electron microscopy characterization of nanomaterials in foods

Electron microscopy is a recognized standard tool for nanomaterial characterization, and recommended by the European Food Safety Authority for the size measurement of nanomaterials in food. Despite this, little data have been published assessing the reliability of the method, especially for size measurement of nanomaterials characterized by a broad size distribution and/or added to food matrices. This study is a thorough investigation of the measurement uncertainty when applying electron microscopy for size measurement of engineered nanomaterials in foods. Our results show that the number of measured particles was only a minor source of measurement uncertainty for nanomaterials in food, compared to the combined influence of sampling, sample preparation prior to imaging and the image analysis. The main conclusion is that to improve the measurement reliability, care should be taken to consider replications and matrix removal prior to sample preparation