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| 10 May 2016 Application Notes bring you the latest information about innovative tools and technologies and their applications in the lab. We hope that you will find this service useful and informative and encourage you to sign up for future Updates to ensure that you never miss one! | ||||||||||
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 | | HyVolution—the smart path to confocal super-resolution www.leica-microsystems.com/ > Super-resolution refers to any device or method that can resolve better than the classical Abbe limit. Apart from infinite super-resolution techniques such as STED (stimulated emission depletion) and SMLM (single-molecule localization methods) that can theoretically resolve to any detail, there are also methods for limited super-resolution. Here we present HyVolution by Leica, which merges optical super-resolution and computational super-resolution. The optical part is provided by confocal microscopy, and the computational part by deconvolution. Lateral resolution of 140 nm is demonstrated. HyVolution offers multiple fluorescence recording in truly simultaneous mode.
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 | | Standardizing protein quantification of purified His-tagged proteins using a micro-volume UV/VIS reader with cDrop™ software www.trinean.com > Purification of recombinant proteins from lysates or supernatants is commonly performed by affinity tagging followed by immobilized metal-affinity chromatography (IMAC). Polyhistidine-tagged (PHT) proteins bind with high affinity to Ni-columns and can subsequently be eluted by adding free imidazole. Buffer exchange of theresulting protein fraction to e.g. PBS and subsequent accurate concentration determination are of paramount importance for the success of ensuing bioactivity assays that assess the therapeutic potential. Purification of process-related impurities like imidazole falsify concentration determination and might impact such assays. Thus, determining both the concentration of purified protein and possible residual imidazole content in the final sample ensures correctness of the actual concentration of the purified target protein. Besides this, it canprovide valuable information about the effectiveness of both purification and buffer exchange methods. In this study, PHT Fab fragments produced at MorphoSys were purified on Ni-columns and eluted with a buffer containing 250 mM imidazole. By combining Trinean's micro-volume DropSense™ 96 spectrophotometer andproprietary cDrop software, the presence of imidazole-contamination can be accurately assessed in all steps of the Fab fragment purification process. Furthermore, in contrast to other classic A280 quantitation methods, cDrop allows determining the actual protein concentration more accurately especially at higher residual imidazole concentrations by correcting for imidazole content.
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 | | Real-Time Cell Health Monitoring, Cell-By-Cell, in 2D and 3D in the Far-Red www.biostatus.com > Inflow cytometry and fluorescence microscopy (including high content screening), a reliable estimation of cell viability is important because it is central to assays for apoptosis and in vitro toxicology. Likewise, it is a measure of sample quality and reliable phenotypic analysis. To improve reporting of such cell viability DRAQ7™ was developed. Essentially, this water-soluble probe has identical spectral properties to DRAQ5™ and does not overlap with visible range fluors. Likewise, RNA binding is very weak (undetectable by flow cytometry). However, DRAQ7™ is membrane impermeant, yet rapidly enters "leaky" cells to label nuclear DNA. Therefore, DRAQ7™ can be used as a new far-red reporter of cell viability and conversely cell membrane-permeabilization resulting from damage, apoptosis and necrosis.
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 | | Diagenode® Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage www.diagenode.com/categories/rrbs-service > Reduced representation bisulfite sequencing (RRBS) enables genome-scale DNA methylation analysis in any vertebrate species. The assay benefits from the practical advantages of bisulfite sequencing while avoiding the cost of whole-genome sequencing. The Diagenode Premium RRBS kit makes this technology widely available and provides high coverage (up to 4 million CpGs in human samples). Multiplexing prior to bisulfite conversion allows processing of 96 samples per experiment, enabling studies of large cohorts.
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 | | QPrEST™—isotope-labeled multipeptide standards for quantitative mass spectrometry-based proteomics atlasantibodies.com/#!/learn-more/about-qprest > Mass spectrometry (MS) enables absolute quantification of endogenous proteins by the use of isotope-labeled standards as internal references. QPrEST standards, currently available for >13,000 human proteins, represent a novel class of recombinantly produced heavy isotope-labeled standards that are added early in the quantification workflow. These multipeptide standards contain 50–150 amino acids identical to a human target sequence, resulting in absolute quantification data based on multiple tryptic peptides.
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 | | Reduced-Bias Small RNA Library Preparation with Gel-Free or Low-Input Options www.biooscientific.com > Changes in microRNA (miRNA) expression have been shown to be associated with a variety of normal physiological processes, as well as diseases including cancer. Studies have already shown that miRNAs may provide useful markers for the development of disease diagnostic and prognostic assays. Next Generation Sequencing (NGS) of small RNAs has significantly improved our understanding of gene regulation, as small RNAs have been shown to fine tune or modulate the expression of genes important in tissue maintenance and disease. NGS brings sensitivity, specificity, and the ability to maximize data acquisition and minimize costs by using multiplex strategies to allow many samples to be sequenced simultaneously with small RNA analysis. This strategy has enabled researchers to perform genome-wide and high depth sequencing studies that would not be possible using other technologies. The ability to quickly and efficiently measure or profile these transcripts has great importance in research and clinical applications.
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