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We narrowed to 706 results for: ESC;

Showing: 41 - 60 of 706 results

  1. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    ...Updated May 10, 2021. In the world of fluorescent proteins and their use for imaging cell biology, Michael...searchable format on Addgene’s Michael Davidson Fluorescent Protein Collection webpage. These plasmids are... 300 plasmid backbones spanning a rainbow of fluorescent proteins “It’s a great collection with over 300... the impressively big deposit. “He has every fluorescent protein under the sun, from the standard oldies...of interest isn’t in the ORF collection, the fluorescent proteins are, which makes mixing and matching...possibilities exponentially as far as new targets and fluorescent proteins are concerned.” For more information...fusions, see this blog post from Joachim Goedhart: Fluorescent Proteins 101: GFP Fusion Proteins - Making the...

  2. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...oligomerization of fluorescent proteins is a concern. The propensity of fluorescent proteins to oligomerize...POI is at the C-terminus of the fluorescent protein. I prefer to describe the different parts of the fusion...of the fluorescent protein are relatively close together (figure 1). Where should the fluorescent protein...length, the specific fluorescent protein variant, and where to add the fluorescent tag. These choices will...dynamic range of fluorescent indicators for Ca2+ by circularly permuted yellow fluorescent proteins." Proceedings...Addgene blog Read our fluorescent proteins blog posts Download the fluorescent proteins 101 eBook Read....org Browse the fluorescent protein collection Find empty backbones for fluorescent protein fusions Browse...

  3. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...Brainbow 3.0 and using the same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication)...default, they are not fluorescent. Instead, they incorporate a truncated, non-fluorescent form of YFP as the...is necessary to excise the non-fluorescent YFP and activate fluorescence of the downstream OFP, GFP, or...combination of all of the fluorescent proteins from the injected construct. The fluorescent protein expressed ...upstream of the fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre...Visit our fluorescent proteins topic page Resources on Addgene.org Browse Addgene’s fluorescent protein...introduction of a transgene construct containing fluorescent proteins (XFP) of different colors to label an...

  4. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ... of TagRFP-T fluorescence intensity due to interactions with photoactivatable fluorescent protein Dronpa...soCoChR-expressing system. Taken together, Shemesh et. al. describe a novel system for the precise mapping of neuronal...the single-cell level. Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid...derivatives encoding alternative promoters and/or fluorescent tags are available at Addgene. Shemesh OA, ...operon, the growth of this knockout strain can be rescued by the xanthine produced via complete demethylation... biochemical activity within the cell. Their Fluorescence fLuctuation INcrease by Contact (FLINC)-based...proximity. By monitoring changes in TagRFP-T fluorescence intensity at different time points, the authors...

  5. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...PPIs): Bimolecular Fluorescence Complementation (BiFC). What is bimolecular fluorescence complementation?...BiFC only uses a single fluorescent protein to visualize PPIs. This fluorescent protein (FP) is split into...complete FP results in a fluorescent signal that can be detected by fluorescence microscopy. Figure..., resulting in a fluorescent readout.   Applications for bimolecular fluorescence complementation BiFC...complementation? Bimolecular fluorescence complementation is a method for detecting protein-protein interactions that...that uses split fluorescent proteins tethered to potential protein interaction partners. Unlike FRET, BiFC... into two fragments that individually lack fluorescence. When complementary FP fragments are fused to ...

  6. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds just over 13,000 significant...those 13,000 regions are important for maintaining ESCs. So which of these 13,000 regions are critical? ... CRISPR out the enhancers from step 2. See which ESCs die or differentiate Step 3 reveals which NANOG-...start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus...average, one distinct enhancer disrupted. As our ESCs die and differentiate over time, we periodically...” for our stem cells. Remove this pillar and the ESC dies. Order Lentivirus from Addgene If you’re interested...established with Bowtie2 using the parameters first described in Kearns et al.: bowtie2 -f -x HG19_GENOME --...

  7. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    ... reduced eGFP fluorescence. Cells expressing the library are then sorted by fluorescence-activated cell... to use a suite of fluorescent reporters. Bacteria containing unstable fluorescent proteins under the ...inoculation, no more fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be...Davidson and Roger Tsien played enormous roles in fluorescent protein development by openly sharing their resources... a conference of their choice to share their fluorescent protein based research. This year, we put out...were excited to see the many creative uses of fluorescent proteins. We’re happy to announce the three winners...levels of abundance, the eGFP will be stable and fluoresce. But if the variant is unstable, it will be degraded...

  8. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...cobalamin-binding riboswitches that bind to cobalamin-fused fluorescent dyes; binding of the cobalamin fusion to the ...the riboswitch results in increased fluorescent signal. Prior to the development of these probes, fluorophore...fluorophore binding aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging...are always on the lookout for better, brighter fluorescent proteins (FP) with improved tissue penetrance...colleagues started with the FP mNeptune684 (originally described by Li ZY, et al. PLoS One, 2016) and tested a ...30220597. ACE system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing...deposited a reporter plasmid that allows for rapid fluorescence-based quantification of base editors that utilize...

  9. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...artifacts. If you're using green fluorescent protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional... are described below (See Figure 1).   Figure 1: Each problem with dimerizing fluorescent proteins...11988576. 3. Matz, M.V., et al., Fluorescent proteins from nonbioluminescent Anthozoa species. Nat Biotechnol... means that your POI fused to GFP or another fluorescent protein (FP) could be forming dimers in cells... by FP dimerization.   1. Transmembrane fluorescent protein fusions Transmembrane proteins, i.e. ...energy to the second FP, often decreasing the fluorescent signal from the first FP and increasing the signal... not necessarily monomeric, even if they were described as monomeric in a publication. The gold standard...

  10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...minutes of inactivating the CASANOVA complex, fluorescent CRISPR complexes were observed binding telomeric...30377362. Studying neuronal activity using the fluorescent voltage indicator Archon Article contributed ...team identified Archon1, a novel opsin-based fluorescent voltage indicator. Archon1 can embed itself into...Heckler et al., 2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and Blatt, 2012...adapted cells expressing these plasmids, tgRFP fluorescence localizes to the mitochondria, because cPYP ...

  11. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...derive mice from these engineered ESCs. To do so, the engineered ESCs would be injected into developing...introduce a mutation into a mouse embryonic stem cell (ESC) line, enrich and select for cells that had successfully...cells in the adult mice derived from the engineered ESCs), and chimeric adults mated to produce completely...need for enrichment and selection offered by mouse ESCs. For example, we often screen only 15-25 mice when...and C57BL/6, for which we have germline competent ESC lines. Second, the process is much quicker. It takes...offspring, with 85% success compared to 50% when using ESC (Cohen, 2016). See Our Recent Post for Tips on CRISPR...

  12. Fluorescent Protein Guide

    Type
    Collection
    ... Plasmid Collections Fluorescent Protein Guide Fluorescent Proteins: Plasmids and Resources... of fluorescent proteins. Use this guide to learn more about the many applications of fluorescent proteins...Addgene Addgene Blog: Fluorescent Proteins Luciferase Plasmid Collection Fluorescent Protein Kits Allen ...this guide to learn about the applications of fluorescent proteins and to find the plasmids that are available...find plasmids for your next experiment. Browse Fluorescent Protein Plasmids by Category Empty Backbones ...of your protein of interest by fusing it to a fluorescent protein. FRET Find FRET pairs and standards to...intracellular processes with genetically encoded fluorescent biosensors. Optogenetics Guide | Optogenetics...

  13. Fluorescent Proteins: FRET

    Type
    Collection
    ... Plasmid Collections Fluorescent Proteins FRET Fluorescent Proteins: FRET FRET Pairs FRET Calibration...Luciferase can be paired with a fluorescent protein acceptor for Bioluminescence Resonance Energy Transfer ...Calibration Standards More Resources Förster (or Fluorescence) Resonance Energy Transfer (FRET) is a process...binding or conformational changes. Organic dyes, fluorescent proteins, and other molecules have been demonstrated...experiments. FRET efficiency can be measured using fluorescence microscopy or spectroscopy by exciting the donor...emission of the acceptor. Alternatively, the fluorescence lifetime (the time between excitation of the...a photon) of the donor can be measured using fluorescence lifetime imaging microscopy (FLIM) or FLIM-FRET...

  14. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ... Plasmid Collections Fluorescent Proteins Fluorescent Biosensors Fluorescent Proteins: Biosensors Metal... AAV Biosensors Fluorescent biosensors are proteins designed so that their fluorescent properties are ...signal of a circularly permuted fluorescent protein (cpFP) or fluorescence resonance energy transfer (FRET... Green fluorescence indicator of K+ for optical imaging (GINKO) Genetically encoded fluorescent indicators...AMP) Yellow fluorescent cAMP indicator (Flamindo2) Genetically-encoded yellow fluorescent cAMP indicator...cyclic AMP) Red fluorescent protein-based cAMP indicator (Pink Flamindo) Red fluorescent protein-based ...SiRI) with GFP-based fluorescent reporter cAMPr Spatial Multiplexing of Fluorescent Reporters for Imaging...

  15. Michael Davidson Fluorescent Protein Collection

    Type
    Collection
    ...Plasmid Collections Fluorescent Proteins Michael Davidson Fluorescent Protein Collection Michael Davidson...Empty Backbones Plasmid Description Industry Return to top Blue Plasmid Description Industry Return to top...top Cyan Plasmid Description Industry Return to top Green Plasmid Description Industry Return to top mEmerald... mEmerald Plasmid Description Industry Return to top Yellow Plasmid Description Industry Return to top...top Orange Plasmid Description Industry Return to top Red Plasmid Description Industry Return to top Photoactivatable...Find fluorescent protein fusion constructs and cloning backbones from the Michael Davidson Lab. Plasmid...Davidson Fluorescent Protein Collection Empty Backbones Blue Cyan Green mEmerald Yellow Orange Red Photoactivatable...

  16. Fluorescent Protein Guide: In Vivo Imaging

    Type
    Collection
    ... Plasmid Collections Fluorescent Proteins In Vivo Imaging Fluorescent Proteins...photoactivatable fluorescent proteins. Plasmid...organs and whole mammals. Genetically encoded fluorescent proteins most suitable for stable imaging of ...undesirable background signals. Many near-infrared fluorescent proteins for in vivo imaging are derived from... photoreceptors (BphPs). Spectrally distinct fluorescent iRFP variants have been shown to have high brightness...of the original iRFP has resulted in a split fluorescence complementation probe used for detecting protein-protein...

  17. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ... Plasmid Collections Fluorescent Proteins Subcellular Localization Fluorescent Proteins: Subcellular ... protein to a red fluorescent protein (find plasmids for cloning in our Fluorescent Proteins Empty Backbones...Collection Michael Davidson Fluorescent Protein Collection Addgene Blog: Fluorescent Proteins Do you have suggestions...Find plasmids encoding fluorescent fusion proteins with known subcellular localization patterns for colocalization...localization. One of the most common is to use fluorescence microscopy to compare the localization of your...trafficking. Addgene has assembled a collection of fluorescent fusion proteins that are targeted to various ... should fuse your protein of interest with a fluorescent protein that has a different emission wavelength...

  18. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...Return to top Fluorescent Timers Fluorescent protein variants that change their fluorescence from the blue... Plasmid Collections Fluorescent Proteins Empty Backbones Fluorescent Proteins: Empty Backbones ...Maturation is the time for fluorescence to reach half its maximal value. Structure describes the oligomeric state...Collection Michael Davidson Fluorescent Protein Collection Botman-Teusink Yeast Fluorescent Protein Collection ...Addgene Blog: Which Fluorescent Protein Should I Use? Addgene Blog: Choosing Your Fluorescent Proteins for Multi-Color...Explore empty plasmid backbones with different fluorescent tags to create fusion proteins with your gene...Resources The following tables list many popular fluorescent proteins and empty backbones for expression and...

  19. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...efficiency of AAV mediated genome editing in human ESCs and iPSCs from ~0.2% to ~1.5%. In more recent work...combining the AAV and CRISPR-based approaches is described in several recent reviews (Howes and Schofield...

  20. Immunocytochemistry

    Type
    Protocol
    ...This protocol describes the basic steps for fixing and staining cells in culture with a primary antibody...antibody against a target protein and a fluorescent secondary antibody....antibodies to detect antigens in cells. Here we describe the basic steps for fixing and labeling cells ...primary antibody against a target protein and a fluorescent secondary antibody. This protocol outlines the... tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10 µL single channel pipette 2...500 µL PBS on a rocking platform. Dilute the fluorescently-labeled secondary antibody to the desired concentration...generally ranges from 1-10 µg/mL. Add 500 µL fluorescently-labeled secondary antibody to the wells and ...
Showing: 41 - 60 of 706 results