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We narrowed to 478 results for: FLU

Showing: 121 - 150 of 478 results
  1. Getting to Know Expansion Microscopy

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    Blog Post
    ...Stiles MA (2022) Improved fluorescent signal in expansion microscopy using fluorescent Fab fragment secondary...101: Introduction to Immunofluorescence  Antibodies 101: Multiplex Immunofluorescence Antibodies 101: Affinity...workings of the synapse by conventional light or fluorescence microscopy (Dani, et al., 2010). How do you ...the resolution barrier while still utilizing fluorescence microscopy, is another option. However, most...understand the ultrastructural relationships that influence everything from cell division to memory, but don...antibodies, chemical dyes, and genetically encoded fluorescent proteins (Gambarotto, et al. 2019; Tillberg, ... of cells and tissues labeled using standard fluorescent proteins and antibodies. Nat Biotechnol 34:987...
  2. Brain Initiative Collection

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    Collection
    ...genome encoding Cre-dependent expression of the fluorescent protein EYFP from the CAG promoter Viviana Gradinaru...An AAV genome that ubiquitously expresses the fluorescent protein eYFP from the CAG promoter Viviana Gradinaru...Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and targeted to the neuronal soma and proximal... AAV production plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent expression Edward...
  3. Using AAV for Neuronal Tracing

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    Blog Post
    ...agglutinin WGA, Schwab et al., 1978) or fluorescent molecules such as Fluoro-Gold (Schmued and Fallon, 1986) ...Schmued L.C., and Fallon J.H. (1986). Fluoro-Gold: a new fluorescent retrograde axonal tracer with numerous...either produce a colorimetric product or are fluorescent. While dyes usually spread unidirectionally, ...biotinylated dextran amines (Veenman et al., 1992) or fluorescent molecules like rhodamine-isothiocyanate (RITC...high and neuron-specific transgene expression. Fluorescent proteins are widely used for tracing and their...with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection into a conditional mutant mouse...., 2017) that express one of a few different fluorescent proteins (red, green, blue). If a researcher ...
  4. Evolution of Lab Techniques

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    Blog Post
    ...labeled probes or by the presence of fluorescence for fluorescently labeled probes. Colorimetric and luminescence...nucleotides were replaced with fluorescently labeled ones allowing for fluorometric-based detection. This increased...synthesized by electronically recording fluorescence activity as fluorescently labeled nucleotides are incorporated...through hybridization of a radioactively or fluorescently labeled probe. If the DNA is complementary to...quantitative PCR as an alternative to Southern blot or fluorescence in situ hybridization for detection of gene ...DNA by restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy of Sciences...
  5. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

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    ...elegans even easier - now, one can generate a fluorescent protein fusion, transcriptional reporter, and...self-excising cassette (SEC). To make it possible fluorescently label proteins in C. elegans, they inserted ...inserted the SEC into a synthetic intron between a fluorescent protein (FP) and a 3xFLAG tag (see construct schematic...located in this synthetic intron, and only the fluorescent protein and 3xFLAG-tag sequences are inserted...serve as the repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template...the Roller phenotype. These animals contain a fluorescent protein fusion for the targeted gene. After initial...Dickinson et al. (2015) illustrating the steps to fluorescently tag a C. elegans protein. The his-72 locus is...
  6. Visualizing Genomic Loci with CRISPR-Sirius

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    Blog Post
    ...al., 2015). To do so, they fused a different fluorescent protein to various Cas9 variants, SpCas9, NmCas9...PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to the genomic loci...to the small molecule, DFHBI-1T to induce its fluorescence. They found that aptamer insertion in the gRNA...8XMS2 which will recruit the HaloTag with red fluorescent ligands to the loci of interest. Dual color detection...sgRNA-Sirius-4X(MS2-PP7)) that result in yellow fluorescence at the loci of interest, the team could visualize...4X(MS2-PP7) showing recruitment of different fluorescent proteins. (B) CRISPR-Sirius used to visualize...loci, what makes CRISPR-Sirius unique? Unlike fluorescent in situ hybridization, CRISPR-based visualization...
  7. A Control for All Seasons

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    ...Burry 2011). Autofluorescence Endogenous fluorescence of the sample, or autofluorescence, also interferes...without any antibodies to determine the level of autofluorescence in the sample. D, when using multiple primary...than the background noise (Waters 2009). If the fluorescence from your target is extremely bright, then the... the noise likely will not interfere. If the fluorescence from your target is weak, then it is best to...and labels omitted. Any signal remaining is autofluorescence and should be considered in the interpretation...controls are single stained samples for each fluorophore and are critical to analyze multiplex flow cytometry...Jennifer C. “Accuracy and Precision in Quantitative Fluorescence Microscopy.” Journal of Cell Biology, vol. 185...
  8. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    ... for awards to early career scientists using fluorescent proteins in their research. These awards help...roles in the development and dissemination of fluorescent protein tools. This year, we’ve made some changes...-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study glutamate, a signaling...disease,” says Aggarwal. This biosensor changes fluorescence based on the release of glutamate from neurons...to the roGFP2 cysteine thiols and change its fluorescence properties. The biosensor can also be used along... GCaMP7, measuring calcium signaling related fluorescence during morphine administration and withdrawal...agonist inhibits the neuron (via hM4D). She fused fluorescent protein tags to the receptors to monitor their...
  9. Antibodies 101: Conjugation

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    Blog Post
    ...together. Common conjugates in the lab Fluorophores Fluorophores allow you to detect your antibodies using...techniques like fluorescence microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available...tool for comparing different fluorophores. We also touched on fluorophores as antibody conjugates in an...sure to check the spectral properties of the fluorophores you are considering to make sure that they’ll...equally, if not more, common conjugate class than fluorophores are enzymes. The most common enzyme conjugates...
  10. Lighting Up Cell Signaling with Photoswitchable Kinases

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    ... the active site. Dronpa also produces green fluorescence when exposed to 500 nm light.  Under 400 nm ... kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR is a ...active, it phosphorylates the ERK KTR and red fluorescence localizes to the cytoplasm. When MEK1 and ERK...inactive, the ERK KTR isn’t phosphorylated and red fluorescence can be found in the nucleus. For testing kinase...Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox... KTR reporter plasmids Find photoactivatable fluorescent proteins in empty backbones in the Michael Davidson...Photoswitchable Fluorescent Proteins ...
  11. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    ...and/or fusing your gene target sequence to a fluorescent reporter sequence (discussed below). Examples...Non-destructive methods, most popularly with a fluorescent reporter, directly couple silencing of your gene... that contains a target sequence against the fluorescent reporter in an inverted repeat orientation. Therefore...have readily observable phenotypes quantified. Fluorescent reporter-based RNAi constructs have been designed...presence of the RNAi transgene, followed by a fluorescence readout of RNAi activity. As a plant researcher...subpopulation can be easily achieved through fluorescence-activated cell sorting (FACS). This is not possible...visually identified as undergoing RNAi through the fluorescent reporter and manually extracted. This process...
  12. Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart

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    ...improved fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent protein... addition to creating and sharing many great fluorescent protein tools, Professor Goedhart is very active..., characterizes and uses genetically encoded fluorescent probes. You can follow him on twitter: @joachimgoedhart...Additional Resources from the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's Series... Series on Choosing the B(right)est Fluorescent Protein Plasmids 101: Visualizing Subcellular Structures...and Organelles Resources at Addgene.org Find Fluorescent Protein Plasmids Find Plasmids for FRET Find ...
  13. Early Career Researcher Toolbox: Finding Relevant Papers

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    ...as the number of citations highly influenced by this paper. Influence is measured by the number of citations...or are highly influenced by the author. You can also see who an author is most influenced by (other authors...learning to extract important features such as influential citations, images, and key phrases. It also takes...help you quickly identify papers with greater influence, the average number of citations per year and ...authors who they cite the most), and who the author influences (who cites the author the most). Best uses Finding...Results in your feed are ranked based on their influence on the network, or body of literature, as a whole...
  14. Plasmids for Endogenous Gene Tagging in Human Cells

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    ...Institute for Cell Science Plasmid Collection Fluorescent tagging strategy Figure 1: Gene editing... long stretches of DNA on either side of the fluorescent protein sequence that is homologous to the sequence...through homology directed repair To introduce the fluorescent tag to the cells, we used CRISPR/Cas9 to make...will attempt to repair itself without outside influence (non-homologous end joining, or NHEJ), in approximately...cells, and C. elegans. In addition to generating fluorescent protein fusions, plasmids are available for tagging...gene of interest. Endogenous gene tagging with fluorescent proteins as described here enables live cell ...about Non-Homologous End Joining Check Out Our Fluorescent Protein Posts Resources on Addgene.org Browse...
  15. Illuminating Epigenetics with A FRET Based Biosensor

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    ...either through histone modifications, which can influence the transcription of genes by RNA polymerase II...techniques include chromatin immunoprecipitation, fluorescent in-situ hybridization, methylation-sensitive ...facilitated by the flexible linker) that brings the fluorescent units of the FRET pair together and causes an...signal. Subsequent demethylation separates the fluorescent units and so lowers the FRET signal back again...Change in paternal grandmothers early food supply influenced cardiovascular mortality of the female grandchildren...Jao, and Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in living cells...the Ting Lab plasmids at Addgene Browse More Fluorescent Protein Biosensors Read Our Tips for Using FRET...
  16. Polymerase Chain Reaction Overview and Applications

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    Blog Post
    ...probe based fluorescent dye that intercalates with any dsDNA and the use of a fluorometer feature built... thermocycler to measure that fluorescent output. With this fluorescent dye, the concentration of the ...which measures the fluorescence amplitude of each droplet. The fraction of fluorescent PCR-positive droplets...reaction cycles is continuously detected via a fluorescent signal. The signal increases proportionally to...concentration of the starting template DNA the fluorescent signal throughout the reaction is compared to...
  17. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...comparing the threshold fluorescence of the viral DNA to the threshold fluorescence of a standard - a set...plate and measures the fluorescence amplitude of each one. Droplets that fluoresce contain the amplified...been read, the software outputs an image of the fluorescence amplitude measured for each droplet in each ...droplet readers have a few channels for detecting fluorescence so that it is possible to measure the concentration...After additional rounds of amplification, your fluorescence amplitude will increase, leading to greater ...al. "A systematic investigation of parameters influencing droplet rain in the Listeria monocytogenes prfA...
  18. Tips for CRISPR Gene Editing in Mice

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    ...incubation, you assess the level of fluorescence in your cells under a fluorescence microscope. The higher the ... homology dependent repair by enhanced green fluorescent protein (EGFP) reconstitution. (a) Scheme of ...containing sgRNA sequences into HEK293T cells. The fluorescence intensity was classified into four groups (4...darker than control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted...your target is inserted, disrupting the EGFP fluorescent signal, for details on this plasmid, see Mashiko...the amount of fluorescence, the better that gRNA is at causing a double stranded break at the target site...screening as little as 8-10 pups. The level of fluorescence you saw in the validation assay should give ...
  19. Our Most Popular Blog Posts from 2019

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    ...CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These resources ...Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins... tools used in cloning. 8. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi by Jennifer...and mWasabi and find plasmids containing these fluorescent proteins at Addgene. 7. Designing Your Chalk...chalk talk, and tips for practicing.  6. The Fluorescent Vegetables in Aptamer Soup by Aliyah Weinstein...aptamers, which are nucleic acid sequences that fluoresce upon binding a small molecule, are often named...making scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the...
  20. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

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    ...when paired with appropriate fluorescent optics. Existing red fluorescent voltage indicators, however,...31937940. Detection of neural activity using a red fluorescent genetically encoded voltage indicator Article...voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight...manipulations when combined with existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins...zebrabow. In this system, cells express various fluorescent proteins controlled by Cre recombination. Each...variety of colors depending on the combination of fluorescent proteins expressed. CreLite-mediated recombination...
  21. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

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    ...Resources Cre-lox Resource Guide Fluorescent Protein Resource Guide Fluorescent Protein Blog Posts ...constructs similarly to GFP, but commonly used red fluorescent proteins dsRed, mCherry, and TdTomato do not ...GFP and derivatives GFP and YFP, but not red fluorescent proteins, as seen with T-DDOGs. Like T-DDOGs...utliized as scaffolds, including the set of red fluorescent proteins. The T-DOGGs and Cre-DOG are available... C.Y., et al. “A nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene ... Caussinus, E., Kanca, O., and Affolter, M. “Fluorescent fusion protein knockout mediated by anti-GFP ...
  22. A Primer on Optogenetics: Introduction and Opsin Delivery

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    ...co-localization immunofluorescence, cell-type specificity. Viruses that only express the fluorescent proteins...chloride ion pumps, activation of which results in efflux of chloride ions, subsequent hyperpolarization,...unlike HR, this is achieved through hydrogen ion efflux; evidence suggests that Arch may more effectively...Thankfully the Deisseroth constructs coexpress a fluorescent protein, which allows for easy identification...general rule of thumb, if you can detect a strong fluorescent signal (without antibody manipulation), then ... vectors for anterograde axonal tracing with fluorescent proteins in nontransgenic and cre driver mice...optogenetics plasmids! Find AAV resources! Find fluorescent protein resources!    ...
  23. Technical Design of a Western Blot

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    ...amount of protein you are expecting.  Fluorescent proteins Fluorescent proteins, while more expensive, allow...typically rely on primary antibodies conjugated to fluorescent proteins, instead of the more common chemiluminescent...you to optimize for smaller proteins or lower fluorescent backgrounds. However, PVDF does add a few (short... of reporters: chemiluminescence enzymes and fluorescent proteins. Chemiluminescence enzymes are cheap...important if you want to quantify your blots). Fluorescent proteins are stable, so blots can be stored for...magnitude, which includes a wide dynamic range for fluorescent proteins. They do tend to have increased background...usually more expensive Nitrocellulose or low-fluorescence PVDF membranes Increased fragility (nitrocellulose...
  24. Luciferase Plasmid Collection

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    ...donor and HaloTag labeled with a fluorophore as the acceptor. LumiFluors: EGFP-NanoLuc ( GpNLuc ) and LSSmOrange-NanoLuc...Nanoluc®. CalFlux VTN : An intracellular calcium sensor created by fusing NanoLuc® and Venus fluorophore to ...Resources Promega Plasmid Collection Fluorescent Proteins Guide Fluorescent Biosensor Plasmids Blog: Luminescence...Addgene's collection include: Firefly luciferase (Fluc) Renilla luciferase (Rluc) Gaussia luciferase (Gluc...luciferase expression Stephen Tapscott 83281 pAAV-CAG-FLuc Firefly CAG AAV expression of firefly luciferase...luciferase Mark Kay 105532 pAAV.CMV.ffLuciferase.SV40 Firefly CMV AAV expression of firefly luciferase James Wilson... Wilson 101156 T7-CMVtrans-FFLuc-polyA Firefly T7 Expression of firefly luciferase from a cell-free mammalian...
  25. Tag Your Favorite Yeast Genes with Ease

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    ...direct fusion of fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes ...several ready-to-use, modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection...of course—and start tagging! Yeast-optimized fluorophores for imaging Lee S, et al. (2013) PLoS ...marker. Lee et al.(1) assessed many of these fluorescent tags in S. cerevisiae, looking at their performance...requirements. Select from these yeast-optimized fluorophore tagging vectors for your single or multi-color...
  26. Genetic Code Expansion

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    ... Mehl 197576 pAcBac1-Mb-Fluoro-Phe B5 Pyl Fluoro-Phe B5 M. barkeri fluorinated phenylalanine derivatives... Mehl 197577 pAcBac1-Mb-Fluoro-Phe D6 Pyl Fluoro-Phe D6 M. barkeri fluorinated phenylalanine derivatives...5FTrp)RS Methanogenic archaeon 4-Fluoro-L-Tryptophan (4FTrp), 5-Fluoro-L-Tryptophan (5FTrp) Bacterial TAG...7FTrp)RS Methanogenic archaeon 6-Fluoro-L-Tryptophan (6FTrp), 7-Fluoro-L-Tryptophan (7FTrp) Bacterial TAG...TAG Chang Liu 85484 pDule-tfmF A65V S158A tri-fluoromethyl-phenylalanine synthetase M. jannaschii family...Wenshe Liu 137908 PylRS-AS PylRS M. mazei fluorophenylalanine derivatives Bacterial TAG Wenshe Liu 137976...163916 pXPR-SF61 SF5Phe tRNA synthetase para-Pentafluorosulfanyl-Phenylalanine Bacterial TAG Thomas Huber ...
  27. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

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    ... vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent protein biosensors...developed in vivo bacterial reporters that use fluorophores and luciferases to track bacteria in real time... reporters Limited Depth of Signal/Imaging: Fluorescent reporters are not as well suited for imaging ...context/spatial localization: In vivo imaging with fluorescent or bioluminescent reporters usually only detects...tools The luciferase reporter gene The green fluorescent protein reporter gene Resources on Addgene.org...
  28. Understanding GPCR Signaling Using TRUPATH

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    ...FRET), in which a donor fluorophore is used to excite an acceptor fluorophore. The difference for BRET...blog Browse all of our fluorescent protein blog posts Find other fluorescent biosensors Resources on...transfer is between a luminescent donor and a fluorescent acceptor. TRUPATH uses an updated version of ...on Addgene.org Browse the catalog of fluorescent protein plasmids Find biosensors for your experiments...
  29. RANbodies: Reporter Nanobody Fusions

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    ...and instead are conjugated to reporters (i.e. fluorescent dyes, HRP, etc. ) after purification. Antibodies...signal amplification occurs when HRP converts a fluorescently labeled tyramide molecule into a highly reactive...linked to residues within close proximity thus fluorescently labeling them. HRP RANbody staining results ... ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to lower background...and tissue sections Amplifying weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution... from Penn State Need help picking the right fluorescence microscopy technique to use? Check out this ...
  30. Hot Plasmids and Viral Preps - January 2021

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    ... in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the...32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin Ng High-throughput ...High-throughput screening methods, such as fluorescence-activated cell sorting (FACS) helps researchers sift through...million cell types expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP ...Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand break repair assay...the introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim...
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