... Plasmid Collections Immunology Research Immunology Research Plasmids and Resources You...Addgene’s immunology collection contains plasmids for genes involved in innate and adaptive immune signaling...secondary immune response. Browse Plasmids for Immunology Research Genes have been grouped into tables ... ImmPort, toward repurposing of open access immunological assay data for translational and clinical research...

...of 15% iodixanol step 5 mL of 25% iodixanol step 5 mL of 40% iodixanol step 3 mL of 60% iodixanol step ...and loading of the iodixanol gradient: 15% iodixanol step: mix 4.5 mL of 60% iodixanol and 13.5 mL of 1 ...buffer 25% iodixanol step: mix 5 mL of 60% iodixanol and 7 mL of 1X PBS-MK buffer and 30 μL of phenol red 40%... 40% iodixanol step: mix 6.7 mL of 60% iodixanol and 3.3 mL of 1X PBS-MK buffer 60% iodixanol step: mix... Purification by Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient Ultracentrifugation...medicine, iodixanol can be used as an intravenous isomolar contrast agent. In research, iodixanol is used...On average, iodixanol-purified AAV preparations contain ~20% empty particles. 60% iodixanol solution is...

...qPCR Using SYBR Green Technology AAV Titration by qPCR Using SYBR Green Technology You may also like......particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s AAV Titration protocol. Protocols...genome-containing particles of an AAV prep using SYBR green technology. This protocol was validated using an internal...

...when we share? NanoLuc® Luciferase-based tools are fantastic examples of technologies that can accelerate...research materials and information. Many of our technology platforms like HaloTag® Fusion Protein, codon-optimized...codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are available from the repository. ...accelerate your research. The creation of NanoLuc® Luciferase from a rather dull 19kDa Oplophorus luciferase...luciferase by Promega’s Advanced Technologies group as well as evolution of the coelenterzine substrate to the...I’ll cover two great applications of the NLuc technology. The first couples NLuc to fluorescent proteins...fluorescent proteins to make better biosensors. NanoLuc®-fluorescent protein fusions for imaging Fluorescent...

...dramatic spread of the Cas9 technology: "The rapid adoption of the Cas9 technology was also greatly accelerated...you think these technologies are going? What is the next hot molecular biology technology? Have you deposited...unique position to observe and quantify how new technologies are being disseminated through the scientific... plasmid sharing In a recent review on CRISPR technology, for example, the authors Patrick Hsu, Eric Lander...

...toolkit based on the Multisite Gateway® technology from Life Technologies. The MultiSite Gateway Kit is a...continually impressed with the amazing plasmid technologies developed by our community of depositors. With...plasmids 62887 & 62888) Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149... system. The ease and flexibility of the LACE technology makes this system widely accessible for many ...imaging and inducible gene expression Brilliant technologies adopt basic biological concepts and employ them... the Weissman lab have teamed up and advanced technology in the fields of single molecule imaging and ...possibility of dimer formation. Prosser et al., Biotechnol Lett. 2015. 37:383-389. Toolkit for constructing...

...Addgene which allow scientists to use CRISPR-Cas technology to perform genome-wide knockout screens. These... Looking for background information on CRISPR technology? See our improved CRISPR-Cas Guide. Wang et...new tool for peptide-peptide locking. The new technology is known as SpyLigase and is a protein domain...E690-7. For further reading about SpyLigase technology, read the Addgene interview with Mark Howarth... and his team at the Institute of Science and Technology Austria, decided to take a novel approach to ...doi: 10.1038/nmeth.3179. Martell et al., Nat Biotechnol 2012 Nov;30(11):1143-8.  A new tool for CRISPR...20;2(9):537-41. DREADD-based chemogenetic technologies After several years of distributing his...

... distributed plasmid technologies pSV-PGC1 - Ok, it's not actually a technology, but pSV-PGC1 was Addgene's...institutions. This has given us a unique window into technology trends in the life sciences. In this post, we'll...give you an inside look at the Top 10 plasmid technologies distributed through Addgene over our first 10...genetically encoded calcium indicators were the hot technology of the year in our collection. CRISPR/Cas9 - ...Year of the CRISPR." The newest genome editing technology has been used and modified by so many labs and...

...the limited technologies available. Over the past few years, however, remarkable technological advances ...and Jinfang Zhu. "Immunologic applications of conditional gene modification technology in the mouse." Current...revolution in modern neuroscience, an era in which technologies like FLEx and optogenetics meet, allowing scientists...mammalian neurons and alter synaptic events: a new technology was born, and the term “optogenetics” was coined... able to distribute materials containing FLEx technology. Read the genOway press release for more information...Deisseroth, Karl, et al. "Next-generation optical technologies for illuminating genetically targeted brain ...at the cellular level in the mouse." Nature biotechnology 21.5 (2003): 562. PubMed PMID: 12665802. SSharma...

...-saturated phenol-chloroform Chloroform 100% ethanol or isopropanol 90% ethanol 70% ethanol TE buffer ...plasmid-containing supernatant. Add either ethanol or isopropanol to precipitate the plasmid DNA. Either ... Optional: Perform phenol-chloroform extraction - see protocol below . Note: Phenol-chloroform extraction...When phenol is mixed with the aqueous solution containing DNA, proteins will move into the phenol phase... either 700 μL of cold 100% ethanol or 350 μL room temperature isopropanol to the solution to precipitate...Protocol: Phenol-Chloroform Extraction of DNA Samples Add an equal volume of TE-saturated phenol-chloroform.... Note: Phenol-chloroform is a hazardous waste - DO NOT pour down sink. Top Protocol: Ethanol Precipitation...

...Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free water...). You can use either Isopropanol or Lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...). You can use either Isopropanol or lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...using an all-in-one acid guanidinium thiocyanate-phenol solution such as TRIzol®. For the precipitation...precipitation step, two options are also included: using Isopropanol or Lithium Chloride. This protocol was adapted...vol) N-laurosylsarcosine (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as TRI Reagent...decontamination solution like RNase AWAY® or RNaseZAP® Isopropanol (for precipitation step, Option A) 7.5 M Lithium...

...Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning...Taipale 87075 pLenti6.2-ccdB-Nanoluc NanoLuc® Creation of C-terminal Nanoluc fusions using Gateway cloning... Taipale 87078 pLenti6.2-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning...Oskar Laur 87070 pcDNA3.1-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning...Proteins Blog: Luciferase 101 Blog: Technologies Enabled by NanoLuc® Luciferase Blog: Luminescent Imaging...interactions. NanoLuc® complementation : Protein-protein assays based on a split NanoLuc® have been used...141285 pGWB401NL3F10H NanoLuc® Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using ...

...15 min in 20% ethanol in deionized water. To prepare 20% ethanol, dilute 2 mL of ethanol into 8 mL of ...Microcentrifuge tubes BCA Assay, Thermo Fisher 23227 β-mercaptoethanol 4X protein loading buffer Precast SDS-PAGE ...-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack, Thermo Fisher IB24002 ...sample for loading as follows: Add 10% v/v β-mercaptoethanol to the 4X protein loading buffer. Dilute 4X...

...egularly I nterspaced S hort P alindromic R epeat) technologies. Before CRISPR, genome engineering approaches...CRISPR one of the most scalable genome editing technologies, making CRISPR perfect for genome-wide screens...population, you will look for relevant hits using NGS technology. For more detail on using CRISPR libraries for...identified by in vivo screening in yeast. Nature Biotechnology , 36 (3), 265–271. PMID: 29431739 Chen, J. S...CRISPR-Cas nucleases in human cells. Nature Biotechnology , 31 (9), 822–826. PMID: 23792628 Fu, Y., Sander...specificity using truncated guide RNAs. Nature Biotechnology , 32 (3), 279–284. PMID: 24463574 Gao, S., Wang...specificity of RNA-guided Cas9 nucleases. Nature Biotechnology , 31 (9), 827–832. PMID: 23873081 Hu, J. H.,...

...TALEN technology and using TALEN plasmids... TALENs TALEN Guide Addgene's Guide to TALEN Technologies You ...'s December 2011 Newsletter Overview of TALEN Technology Imagine being able to manipulate a specific region..., zinc finger (ZF) arrays have been the go-to technology for targeting enzymes and other useful protein.... With the template for studying this type of technology already laid by the zinc finger community ( see...Addgene. This kit, first described in Nature Biotechnology in August, uses a serial ligation protocol to...published a paper on TAL effectors in Nature Biotechnology early in 2011 and they described their kit for...

...this technology is used for targeted neuronal manipulation. Practical tips for using the technology in ...Education and Development (AAVED) Series AAV Technologies and Methods for Targeted Neuronal Manipulation...the guide. We hope that this guide will improve technology adoption and scientific outcomes. Event Details...Cepko 2:45 PM Break 3:00 PM (Group Discussion) Technologies and Resources Missing in the Field Kenton Woodard...presentation is to: Give a general overview of the technology and how it can be used for targeted neuronal ...focused on: Potential combination with other technologies to achieve different experimental goals. How... manipulation? Best practices for using this techonology in the lab (main issues/factors that researchers...

... genome-wide screening. CRISPR Technological Advances CRISPR technology development has occurred at a ...various biological and translational technologies. Biotechnology companies are exploring therapeutic applications...applications, eclipsing past genome engineering technologies like zinc finger nucleases (ZFNs) and TALENs...Scientists have also developed CRISPR editing technologies that do not rely on NHEJ or HDR: Base editors... it’s clear that CRISPR is a truly disruptive technology for basic research. The beauty of CRISPR is that...nuclease specificity using truncated guide RNAs. Nat Biotechnol . 32(3):279-84. PMID: 24463574 Fujita T, Fujii...and engineered sgRNAs using CRISPRainbow. Nat Biotechnol . . PMID: 27088723 Makarova KS, Wolf YI, Alkhnbashi...

...Zebrafish ( Sander et al., Nat Biotechnol. 2011 ) Rats ( Tesson et al., Nat Biotechnol. 2011 ) Human somatic cells...cells ( Miller et al., Nat Biotechnol. 2011 ; Reyon and Tsai et al., Nat Biotechnol 2012 ) Human pluripotent...Ligation) assembly method (Sander et al., Nat Biotechnol. 2011), the REAL-Fast assembly method (Reyon ...) assembly method (Reyon and Tsai et al., Nat Biotechnol 2012). All three of these platforms assemble ...pluripotent stem cells ( Hockemeyer et al., Nat Biotechnol. 2011 ) TALE transcriptional activators in human cells...

...capacity to give rise to an entire organism. iPSC technology removes the possible ethical concerns related...tumorigenicity, an inherent disadvantage of iPS cell technology. Browse Addgene's collection of Transdifferentiation... lineage conversion of acinar cells. Nature Biotechnology. 2014 Dec;32(12):1223-30. Zhou Astrocytes Dopaminergic...oligodendroglial cells by direct lineage conversion. Nat Biotechnol. 2013 May;31(5):434-9. Wernig Fibroblasts Oligodendrocyte...myelinogenic oligodendrocyte progenitor cells. Nat Biotechnol. 2013 May;31(5):426-33. Tesar Fibroblasts Medium...fibroblasts into antigen-presenting dendritic cells. Sci Immunol. 2018 Dec 7;3(30). pii: 3/30/eaau4292. Pereira ...