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... 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...

... and bioinformatic technologies are used.  Improvements to sequencing technologies and bioinformatic analysis...the sequencing technologies that Addgene uses in the QC process, when these technologies are used, and ...up with the constantly updating technologies. Sequencing technologies used at Addgene Sanger sequencing...Nanopore sequencing technology, bioinformatics and applications. Nature Biotechnology, 39(11), 1348–1365...using an Illumina MiSeq, but that’s not the only technology Addgene uses to ensure accuracy of deposited ...Sanger, was the first widely used DNA sequencing technology. The process relies on “sequencing-by-synthesis... Figure 1: Overview of Sanger sequencing technology. Created with BioRender.com. Sanger sequencing...

...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...

...Bryan Welm 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...Javier Alcudia 113442 pcDNA3.1 NL NanoLuc® CMV Mammalian expression of Nanoluc with a N-terminal Myc tag Erich...Rudolf Jaenisch 113450 pLenti NL NanoLuc® Ubc Lentiviral expression of Nanoluc with a N-terminal Myc tag Erich...the acceptor. LumiFluors: EGFP-NanoLuc ( GpNLuc ) and LSSmOrange-NanoLuc ( OgNLuc ) fusions for BRET-based...

...about NanoLuc technology (Link opens in a new window) from Promega Read about HaloTag technology (Link...Industry Plasmid Technology Overview This collection includes specialized Promega technologies that support...support a range of applications. The technology includes NanoLuc, HaloTag, SmBiT, LgBiT, HiBiT, and more. Luciferase...high-throughput screening in drug discovery. NanoLuc Fusions NanoLuc (Link opens in a new window) is a 19.1 ...and cell-free systems. NanoBiT Assays NanoLuc Binary Technology (NanoBiT) (Link opens in a new window)...analysis and fusion vectors containing tags such as NanoLuc, HaloTag, SmBiT, and LgBiT, useful for detecting... Ordering & Availability All Plasmids Plasmid Technology Overview Additional Resources (Link opens in ...

...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...

... 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...

...-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...

...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 ...

...trans-activating crRNA (tracrRNA; Hsu et al ., Nature Biotechnology 2013). We also have an older system using two...express up to three sgRNAs (Swiech et al., Nature Biotechnology 2014). Full references are below. Available ...mouse CRISPR-Cas9 is a versatile genome editing technology for studying the function of genetic elements...Cradick TJ, Marraffini LA, Bao G, Zhang F. Nat Biotechnol. 2013 Sep;31(9):827-32. doi: 10.1038/nbt.2647...Habib N, Li Y, Trombetta J, Sur M, Zhang F. Nat Biotechnol . 2015 Jan;33(1):102-6. doi: 10.1038/nbt.3055...

... promoter in a 'Cre-on' dependent manner. 1 Don Arnold 169128 pAAV-DIO-EF1α-mKate2-PDE4D3-Cat EF1a-driven...viruses infected in postsynaptic cells. 5, 8 Don Arnold 232352 pAAV-DIO-ATLASsnFlp Syn-driven, Cre-dependent...fDIO-Reporter viruses infected in postsynaptic cells. 8 Don Arnold 232353 pAAV-DIO-BACE-HA Syn-driven, Cre-dependent...expressed (1:1) with either ATLAS protein. 8 Don Arnold Don’t See What You’re Looking For? Our Packaged...

...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...

...opens in a new window) based on yeast display technology (Link opens in a new window) and an optimized...cone receptors. Developed using yeast display technology, these antibodies enable high-confidence targeting...synaptic cleft. Developed using yeast display technology, these antibodies provide researchers with powerful...

...England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental Information New England...fixed cells. The plasmids associated with this technology are available at Addgene for distribution to ...NEB's comparison of SNAP-tag, CLIP-tag, and GFP . Technology SNAP-tag SNAP-tag is based on the human O6-alkylguanine-DNA-alkyltransferase...

...monoclonal antibody fragments by phage display. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. Carlos Barbas...and utilisation of recombinant antibodies. BMC Biotechnol. 2006 Dec 7;6:46. John McCafferty CRISPR system...selected from Tomlinson I+J phagemid libraries. Immunol Lett. 2015 Oct;167(2):95-102. Joanna Bereta Sybody...

...units in the oleaginous yeast Y. lipolytica - Rossignol Lab Yeast Gateway kit : 285 vectors for expression...CRISPR Guide for a general introduction to CRISPR technology. Check out our CRISPR reference page for CRISPR...resources Bacterial CRISPR Resources The CRISPR technology was developed from bacterial immune systems, ...

...CRISPR Guide for a general introduction to CRISPR technology or the zebrafish CRISPR resources for a full ...zebrafish resources and developing new lines and technology. European Zebrafish Resource Center (EZRC) - ...- A repository at the Karlsruhe Institute of Technology (KIT) for the distribution of zebrafish lines ...

...encoded calcium indicator for in vivo imaging. Nat Biotechnol. 2021 Mar;39(3):368-377. Vladislav Verkhusha ...pentose and disaccharide accumulation in bacteria. Biotechnol Biofuels. 2008 Jun 3. 1(1):11. Wolf Frommer Maltose...indicator for in vitro and in vivo studies. Nat Biotechnol. 2018 Sep;36(8):726-737. Yulong Li Acetylcholine...imaging of endocannabinoid dynamics in vivo. Nat Biotechnol. 2021 Nov 11. pii: 10.1038/s41587-021-01074-4...measurements of membrane charges in live cells. Nat Biotechnol. 2017 Mar 13. Katharina Gaus Endo/Exocytosis ...

...interest) can be cloned into the plasmid. Garry Nolan 12257 pWPXL 2nd EF-1alpha-driven constitutive transgene...vector system. Expresses gag-pol and rev . Garry Nolan 8455 pCMV-dR8.2 dvpr 2nd Contains gag , pol , rev...recommended for the FELIX (FIV-based) system. Garry Nolan 8454 pCMV-VSV-G VSV-G-expressing envelope plasmid...

... Stem Cell Research page. CRISPR Tools CRISPR technology is a promising therapeutic approach for precise...CRISPR Guide for a general introduction to CRISPR technology or the mammalian CRISPR resources for a full ...Anti-MECP2 (D4F3) rabbit antibody from Cell Signalling Technology Additional Resources (Link opens in a new window...

...CMV Huntington's Arnold Boersma 175061 pQ39-VC HTT Venus, mCherry CMV Huntington's Arnold Boersma 175062...mCherry CMV Huntington's Arnold Boersma 175064 pQ79 HTT mCherry CMV Huntington's Arnold Boersma 175066 pVC-...mCherry CMV ALS Arnold Boersma 175067 pVC-FUS-R521C FUS Venus, mCherry, HA CMV ALS Arnold Boersma 175068...Corporation 237172 EIF2AK2(252-551)-NanoLuc Fusion Vector EIF2AK2 NanoLuc CMV VWM disease Promega Corporation...2A Tau WT WPRE MAPT CBA Parkinson's, FTD Matthew Nolan 138134 pT7NT*-Bri2 113-231 R221E ITM2B NT T7 Familial...175068 pFUS-WT-VC FUS Venus, mCherry, HA CMV ALS Arnold Boersma 175307 pH6HTC_HNRNPA1-Halo HNRNPA1 Halo, His... Arrowsmith 211083 pNLF1-C_EIF2B2:1-351 EIF2B2 NanoLuc CMV VWM, CACH Cheryl Arrowsmith 211112 pBiT3.1-...

...more information on the wide range of CRISPR technologies, visit our CRISPR Guide . There are many different... for CRISPR-Based genome editing. Trends in Biotechnology , 36 (2), 173–185. https://doi.org/10.1016/j.tibtech... analysis of CRISPR–Cas experiments. Nature Biotechnology , 38 (7), 813–823. https://doi.org/10.1038/s41587...

...research and development of engineered zinc finger technology. Consortium members Keith Joung and Daniel Voytas... nuclease fusions Related Genome Engineering Technologies TALEN Engineering Reagents CRISPR Plasmids...

...the lab. Equipment Water Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water ...water. You may use a 10% bleach solution or a 70% ethanol solution to wipe down the inside of the water bath...marker that is also resistant to chemicals such as ethanol. Writing on the tops of the tubes and lids of items...

...produced in mammalian cells AAV Purification by Iodixanol Gradient Ultracentrifugation Protocol Mulitchannel...preparation. AAV Titration by qPCR Using SYBR Green Technology Protocol Agarose Gel Electrophoresis Gel electrophoresis... process when requesting samples from Addgene Technology Transfer How to Order A step-by-step walkthrough...

...use Watch the Video! DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA DNA Quantification...the target of your choice AAV Purification by Iodixanol Gradient Ultracentrifugation Purify adeno-associated...Video! AAV Titration by qPCR Using SYBR Green Technology Use qPCR to measure the physical titer of your...

...Ordering and MTA Information Featured Collections NEW NanoLuc and HaloTag Fusions from Promega COVID-19 SARS-...distribution. Please see the Sample Agreements on our Technology Transfer Information page for more information...

...Production in HEK293T Cells AAV Purification by Iodixanol Gradient Ultracentrifugation AAV Titration by ... ddPCR AAV Titration by qPCR Using SYBR Green Technology Content last reviewed: 10 September 2025 Do you...

...biology, DNA origami, immunology, biochemistry, and more. The foundational technologies enabled by this collection...