A wide range of affinity reagents can be converted to proximity probes by attaching DNA strands. This protocol describes the creation of such proximity probes by covalent coupling of oligoncleotides to antibodies that can be used as either primary or secondary antibodies in assays.

Authors: Ola Söderberg et al
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The following protocol is an example on how to perform a proximity ligation assay such as the ones described in a previous protocol attached to theProximity Ligation Assays group. Sample pretreatment may have to be adjusted for individual pairs of antibodies and for each type of sample. But this protocol provides a good starting point for proximity ligation assays.

Authors: Ola Söderberg et al
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The aim of the protocol is to obtain an enriched fraction of intact mitochondria from the yeast Saccharomyces cerevisiae to perform quantitative determination of the activity of respiratory chain enzymes. All the strains used to perform this protocol are derivatives of W303 (K699: MATa ade2-1 trp1-1 can1-100 leu2-3,112 his3-11,15 ura3-52).

Authors: Stefania Magri et al
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Steroids are powerful signalling molecules regulating a variety of physiological processes such as cell proliferation, cell differentiation, and reproduction. Variations in steroid concentrations may hint at hormonal imbalances or metabolic disorders that allow for a diagnosis of human diseases. The measurement of steroid hormone concentrations is relevant to basic research as well as clinical sci...

Authors: Ferdinand Haller, Cornelia Prehn and Jerzy Adamski
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Microarray analysis and RNA-Seq are powerful tools for investigating differential gene expression in animal models of human genetic disease. Often, these models are loss of function mutations introduced by gene targeting or trapping. One difficulty in such experiments, especially where work on congenital malformations is concerned, is that whole embryo or regional dissections will commonly contain...

Authors: Kelly Lammerts van Bureen and Peter Scambler
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The advances in high throughput techniques for analyzing DNA and RNA have the potential to make a major impact on prevention, diagnosis, prognosis and treatment of many human cancers (1). One of the major challenges is to obtain high-quality DNA and RNA molecules. Although several RNA and DNA isolation methods for frozen tissues have been used (2-6), few efforts have been conducted to optimize th...

Authors: Waleska Kerllen Martins
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This is a quick and robust protocol for isolation of High Molecular Weight genomic DNA from Drosophila embryos. We have used DNA isolated using this protocol for fosmid genomic library production.

Authors: Radoslaw Kamil Ejsmont
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Here, for the first time we have shown that a single receptor, CD40, by virtue of its re-localization within cholesterol-rich detergent-resistant membrane microdomains (DRMs), can assemble a signalosome able to induce pro-inflammatory mediators and immunity to L. major, while the same receptor excluded from DRMs, as occurs in cholesterol depleted or L. major infected cells, signals IL-10 productio...

Authors: Abdur Rub
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Suitable matrices to study metastasis and cancer cell growth in the bone microenvironment have been limited to matrices consisting of specific proteins such as collagen or those secreted by immortalised cell lines. Use of the latter is restrictive since these lines retain characteristics representative of a particular stage of differentiation and often do not have the capacity to differentiate fur...

Authors: Johannes C. Reichert et al
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