Nature Methods Application Notes e-UPDATE: 13 December 2011

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13 DECEMBER 2011  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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FEATURED APPLICATION NOTE "The Simple Western™: a gel-free, blot-free, hands-free Western blotting reinvention www.proteinsimple.com > Western blotting is considered the gold standard for protein detection and characterization. Although improvements to individual aspects of Western methodologies have been developed in recent years, none has integrated the entire process onto a single platform. ProteinSimple™ has developed Simple Western™ assays for protein sizing and quantitative immunodetection as an alternative to traditional Western blot analysis. Assays are performed on Simon™, an instrument that integrates and automates all manual operations associated with Western blotting. PDF

		Duolink-“In-cell Co-IP” for visualization of protein interactions in situ www.olink.com > Duolink® offers a user-friendly solution for studying protein-protein interactions. Duolink utilizes a pair of antibodies that are capable of quantitatively reporting even weak and transient protein-protein interactions in natively expressing cells either as countable, bright fluorescent spots for a standard fluorescence microscope or via chromogenic detection for brightfield microscopes. It offers all the benefits of traditional coimmunoprecipitation (Co-IP) and western blotting but with better quantitative precision and more information about cell-to-cell variability and target localization. It is also amenable to high throughput in 96 or 384 wells. PDF

		EndoLISA®: a novel and reliable method for endotoxin detection www.hyglos.de > A new test for the sensitive detection of endotoxin has been developed, based on a lipopolysaccharide-selective, precoated microplate and a factor C–based detection reagent and presented in a complete kit format. The selective capture of lipopolysaccharide (LPS) is achieved using a phage-derived receptor protein exhibiting high affinity and high specificity for the conserved core region of LPS. After binding of sample-LPS to the microplate as the first stage of the assay, the original sample matrix is washed off, thereby eliminating potentially interfering components. In the second stage of the assay, LPS is detected by factor C in a process whereby the principal receptor of the Limulus amoebocyte coagulation cascade reacts with a fluorescence substrate. The new endotoxin test EndoLISA has a detection range from 0.05 EU/ml up to 500 EU/ml. PDF

		ReliaPrep™ Large Volume HT gDNA Isolation System: gDNA isolation from blood samples www.promega.com > The ReliaPrep™ Large Volume HT gDNA Isolation System enables automated purification of genomic DNA (gDNA) from up to 96 samples of whole blood at a time. The system dramatically increases laboratory throughput while conserving reagents by scaling the purification chemistry automatically to each sample's input volume, providing DNA that is ready for storage or for use in downstream applications without a need to resuspend DNA pellets. PDF

		ESGRO-2i Medium: Inhibitor-Based Serum-free Medium for ES and iPS Cell Culture www.millipore.com > Defined serum-free and feeder-free culture of mouse embryonic stem (mES) cells holds many advantages over the classical serum-containing feeder-dependent culture methods, ranging from decreased lot-to-lot variations to ease of culture. In this study, we explore the use of inhibitor-based, serum-free and feeder-free ESGRO-2i medium (EMD Millipore) for culturing mES cells and induced pluripotent stem cells (iPS cells). PDF

		Expresso® Cloning and Expression Systems: Expressioneering™ Technology streamlines recombinant protein expression< www.lucigen.com/ > The Expresso® Cloning and Expression Systems use Expressioneering™ Technology to dramatically increase the speed and efficiency of target gene cloning and soluble protein expression in Escherichia coli. With Expressioneering, PCR products are cloned instantly and directionally into preprocessed expression vectors without sample cleanup or enzyme treatment. The Expresso Rhamnose System is a single-host system ideal for high throughput, allowing cloning in an effortless afternoon and recombinant protein expression the next day. PDF

		Ribo-Zero Gold Kit: improved RNA-seq results after removal of cytoplasmic and mitochondrial ribosomal RNA www.epibio.com > Ribosomal RNA (rRNA) constitutes the majority (>98%) of total RNA preparations. To avoid wasting sequencing reads, it is necessary to remove this abundant RNA before preparing RNA libraries for deep sequencing. We previously developed Ribo-Zero™ technology for cytoplasmic rRNA removal from intact and degraded RNA. Here, we describe the Ribo-Zero Gold Kit: an improvement to the original Ribo-Zero method that allows removal of mitochondrial as well as cytoplasmic rRNA from total RNA preparations. PDF

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