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We narrowed to 475 results for: FLU

Showing: 121 - 140 of 475 results

  1. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...engineer a wide variety of GFPs with diversity in fluorescence brightness, intensity, excitation and emission...derived from the Timer protein - a mutant of the fluorescent protein dsRed that changes irreversibly its color... and in vivo. Merged green and red channel fluorescence images of C2C12 cells stably expressing pTRE-Tight-MitoTimer...for 2 hours, and imaged on the Keyence BZ-9000 fluorescence microscope 8 hours post-pulse. The distinct ...distinct differences of red-to-green fluorescence in individual mitochondria in the networks marks the differences...overexpression or knock-down, where signaling dynamics are influenced by the relatively long life-cycle of a protein...expression causes the induction of both DsRed fluorescence and GFP shRNA. Mice carrying the R26DsRedR; ...

  2. Getting to Know Expansion Microscopy

    Type
    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...

  3. Using AAV for Neuronal Tracing

    Type
    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

    Type
    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

    Type
    Blog Post
    ...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

    Type
    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

    Type
    Blog Post
    ...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. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ... 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 ...

  9. Antibodies 101: Conjugation

    Type
    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. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ... 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...

  11. Genetic Code Expansion

    Type
    Collection
    ... 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 ...

  12. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...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...

  13. Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart

    Type
    Blog Post
    ...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 ...

  14. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...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. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ...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...

  16. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...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...

  17. Polymerase Chain Reaction Overview and Applications

    Type
    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...

  18. Droplet Digital PCR for AAV Quantitation

    Type
    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...

  19. Luciferase Plasmid Collection

    Type
    Collection
    ...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 ...Constructs Other Highlights You may also like... Fluorescent Proteins Blog: Luciferase 101 Blog: Technologies...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...

  20. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...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 ...
Showing: 121 - 140 of 475 results