We narrowed to 478 results for: FLU

...combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters is a useful...been used to produce complex fluorescent reporter constructs. These fluorescent reporters can be used to ...Plasmids 101: Green Fluorescent Protein Resources on Addgene.org Browse our Fluorescent Protein Collection...circuits or sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging...MXS-chaining was designed to create plasmids for fluorescence imaging or flow cytometry applications in mammalian...cells. The modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation...constructs were created, each containing a unique fluorescent protein with minimal spectral overlap between...

...insertion that disrupts fluorescence due to a frameshift. Restoration of fluorescence can only occur through...mCherry expression. Cells can then be sorted using Fluorescence-activated cell sorting (FACS) and the editing...restoration of a frame-shift in mCherry that restores fluorescence to monitor APOBEC-Cas-9 mediated editing. eGFP...individually mutated three codons in eGFP to eliminate fluorescence. This created three inactivated eGFP reporters...the APOBEC-Cas9 editosome would restore eGFP fluorescence, making it easy to quantify single base editing...mutations in eGFP that results in the restoration of fluorescence. Transient reporter for editing enrichment (...Standage-Beier et al. 2019). TREE is a real-time, fluorescent based identification system for the isolation...

...relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity, ...designed novel SL fluorescent biosensors by integrating a SL-insensitive fluorophore (LSSmOrange) as an...control and a SL-sensitive fluorophore (circularly permuted green fluorescent protein or cpGFP) to the ...thoughtfully designed plasmid components with Fluorescence Activated Cell Sorting (FACS), allowing for .../doi.org/10.1101/2020.05.04.076067    Novel fluorescent enzyme biosensors provide direct, specific and...conformational change of the receptor which modulates the fluorescence intensity of the cpGFP without altering that... authors also showed that the receptor-based fluorescent biosensor has a high sensitivity (50 nM-500nM...

...in this post: Yeast-based protein evolution Fluorescent tools for mammalian cells Nanobody purification...enables tracking of protease activity with a fluorescent protein--no affinity purification required! The...doi.org/10.1038/s41592-019-0665-7 Optimizing fluorescent tools for labeling structures and compartments...mammalian cells By Angela Abitua Genetically encoded fluorescent proteins are often used to label  specific structures...biomolecules within cells. However, sometimes these fluorescent proteins might not localize to the right places...ongoing effort to improve genetically encoded fluorescent markers for labeling specific structures within...described several improved genetically encoded fluorescent markers tagged with mTurquoise2, mNeonGreen, ...

...the B(right)est Fluorescent Protein Additional Resources on addgene.org Fluorescent Proteins: Biosensor...a vital tool in neuroscience research. These fluorescent proteins light up in response to calcium, which...indicator, OCaMP. The challenge: Expanding the fluorescent toolbox Modern neuroscience experiments often...often rely on multiplexing, where multiple fluorescent tools are used simultaneously to monitor or control...various teal, green, yellow, orange, and red fluorophores. The spectra for mOrange2 (scaffold of OCaMP...jGCaMP8) remains the gold standard for green fluorescence, while red indicators jRGECO1a and jRCaMP1a ... OCaMP (Aggarwal et al., 2025), based on the fluorescent protein mOrange2 (Figure 2). Figure 2...

...cyan-excitable red fluorescent protein (mCyRFP1). Derived from cyan-excitable organge fluorescent protein (CyOFP1...as a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy... al., 2007) and includes four promoters, six fluorophores with nonoverlapping emission spectra (N- and...Segment on This New AAV Tool    mCyRFP1: Novel fluorescent protein that enables imaging of two biosensors...to that of CyOFP1, a large stokes shift, long fluorescence lifetime, and a stable monomeric form as determined...mCyRFP1 Podcast Segment New bright monomeric red fluorescent protein - mScarlet! The Dorus Gadella lab has...has recently developed a synthetic red fluorescent protein variant they call mScarlet. Bindels et al used...

...to label loci, such as fluorescence in situ hybridization (FISH) or fluorescently labeled TALEs, CRISPR...applications is a colorful system that uses fluorescently labeled Cas9 to label multiple genomic loci .... dCas9 has also previously been adapted to fluorescently label a sequence matching its gRNA, facilitating...St1Cas9. Each ortholog was fused to a different fluorescent protein to create three colors. The specificity...telomeric sequences and showed that different fluorescently labeled dCas9 isoforms are efficiently directed...distances of 75 and 2 Mbp, with the calculated fluorescent distances correlating with the previously established... al. developed this technique using standard fluorescence microscopy, and are thus limited by its lower...

...cre-dependent version at Addgene! Fluorescent protein and control AAV These fluorescent protein expression vectors...exhibits high contrast between peak fluorescence and resting fluorescence. It is useful for activity imaging...exhibits high contrast between peak fluorescence and resting fluorescence.  It is useful for activity imaging...pGP-AAV-syn-FLEX-jGCaMP7b-WPRE) exhibits the brightest resting fluorescence of the GCaMP7 family of calcium sensors and ...the endoplasmic reticulum, but exhibits larger fluorescent responses to GABA. Find the iGABASnFR and its...the control vectors for experiments encoding a fluorescent-tagged molecular tool.   Find pAAV-FLEX-tdTomato...of densely labeled neurons because background fluorescence from inactive neurons is reduced. 105322 ...

...lab fused a fluorescent protein to the hybrid anti-HA scFvs. The addition of the fluorescent protein allows...to amplify fluorescent signals in in vivo imaging: the 10xHA tag increased the fluorescent signal to noise...protein turnover Download the fluorescent protein eBook Browse our fluorescent protein blog posts Resources...Read our fluorescent protein guide Find plasmids for subcellular localization Find fluorescent protein ... part epitope binding region, all fused to a fluorescent protein. “Frankenbody,” like the infamous frankenstein...scFvs) (Inbar et al., 1972) are fused to mature fluorophores, and can bind and light up epitope tagged proteins...as living neurons, and can be fused to other fluorescent proteins and tags including mCherry, HaloTag ...

...DNA Fluorescence dyes Another way to quantify DNA would be to use fluorescent dyes that fluoresce when...and their corresponding fluorescence. That way, you can compare the fluorescence of your sample against...sequencing. Unlike absorbance-based methods, fluorescence-based methods require a standard curve, a set... have to do the calculation yourself as many fluorometers will calculate your sample concentration for... and you don’t need a spectrophotometer or a fluorometer to quantify DNA this way. First, start by pouring... run, DNA fragments move through a micro- or nanofluidic channel and the sample is separated by electrophoresis...These fragments are quantified over time using a fluorescent dye that intercalates into the DNA such that ...

...protein (the FAP - fluorogen activating protein - dL5** in this case), and 2) a fluorogen (the TAP - targeted...photosensitizer - MG-2I in this case). The fluorogen is not fluorescent until it binds its protein partner thus...Up to Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on Addgene.org...the protein complex complex and the MG-2I-dL5** fluorogen that can be used to ablate cells in culture and...It’s job is to bind MG-2I, a photosensitizer fluorogenic dye.  MG-2I is the TAP portion of the complex...which suggests that it could be used with these fluorescent proteins. He et al also compared MG-2I-dL5** ...head-to-head with KillerRed, a genetically encoded red fluorescent photosensitizer protein that generates ROS when...

...panel.  First, fluorophores vary in brightness. You don’t want a super-bright fluorophore, like PE, on ...assay you can do. Most non-HRP conjugates are fluorophores, which work not by reacting to a chemical, but...then read by a detector panel in the machine. Fluorophores are light sensitive, so antibodies conjugated...before you make your purchase. Picking a single fluorophore for an experiment is easy, but what about those...of interest! Research the brightness of your fluorophores before buying and try to inversely pair brightness...possible.      Fig 2: Try to inversely pair fluorophore strength with protein abundance.    Second...Second, you’ll need to consider spectral overlap. Fluorophores signal are actually quite broad, resulting in...

.... This sensor switches from green fluorescence to a red fluorescent state only by simultaneous (1) user-controlled...Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (... (e.g., calmodulin or troponin C). Their fluorescent readouts are used to monitor calcium concentrations... of the sensor, and (3) subsequent change in fluorescence only occurs at high calcium concentrations. ...reading with other measurements, specifically fluorescent ones? Many of the available biosensors use a ... methods Many calcium reporters use a single fluorescent protein (FP), requiring only a simple measurement...Structure-and mechanism-guided design of single fluorescent protein-based biosensors. Nature chemical biology...

...of two fluorescent proteins with very specific spectral properties that limit which fluorophores can be...genetically encoded fluorescent kinase reporters. A KTR is comprised of a fluorescently-tagged substrate ...flexible swapping of fluorescent proteins and multiplexing is simple when fluorescent proteins with limited...The relative ratio of cytoplasmic vs. nuclear fluorescence serves as a proxy for kinase activity.  Advantages...cytoplasm, nuclear export and import rates can influence KTR translocation. Work from the Covert lab suggests...about photoswitchable kinases Learn more about fluorescent biosensors Learn about using phosphoserine to...Resources on Addgene.org Find FRET plasmids Find fluorescent biosensors ...

... Right Connection Photoactivatable Fluorescent Proteins Fluorescent Protein Timers Return to top Protein...interactions using fluorescence microscopy. FRET relies on the principle that when a donor fluorophore is excited...will be able to detect fluorescence both from the donor and the acceptor fluorophore by only exciting the...ratio Fluorescence (GFP) Escherichia coli Gary Yellen 187836 pVoPo-01 Promoter activity Fluorescence (mCherry...activity Fluorescence (mCherry) Escherichia coli Mattheos Koffas 173481 HC-M ATP Fluorescence (GFPmut2...activity Fluorescence (mRFP1) Escherichia coli Mary Dunlop 124605 pSCM001 Cytoplasmic pH Fluorescence (mCherry...Promoter activity Fluorescence (turbo-365) Mycobacterium sp. Tanya Parish 26156 pMV306hsp+FFluc Promoter activity...

... be visualized using fluorophore-conjugated antibodies (i.e., immunofluorescence), or by a colorimetric... antibody, you can visualize the protein via fluorescence or chemiluminescence, precipitate the protein...protein to be detected by chemiluminescence or fluorescence. Western blots are frequently used to compare... primary antibodies that are conjugated to a fluorophore, to avoid the use of secondary antibodies in ... a mixture of cells. The technique of using fluorescently-labeled antibodies to detect proteins is referred...antibodies to bind to intracellular proteins. The fluorescence signal is detected by the flow cytometer and.... Because the signal comes from the antibody-fluorophore conjugate, and is not expressed by the cell itself...

...about other Fluorescent Biosensors A Practical Approach to Choosing the B(right)est Fluorescent Protein Visit...Visit the Fluorescent Protein Featured Topic Page Resources at Addgene.org Find Fluorescent Protein Biosensors...conformational changes lead to an increase in fluorescence intensity from the inserted GFP. After characterizing...expression levels in bacteria and produces stronger fluorescent signals upon sensing glutamate in mouse and ferret...higher expression levels and produce stronger fluorescent signals upon sensing glutamate using both 1-photon... Biosensors Browse All Fluorescent Proteins Find AAV Tools References Marvin, Jonathan S., et al. “An...An optimized fluorescent probe for visualizing glutamate neurotransmission.” Nature Methods (2013) PubMed...

...generate a fluorescent output, which will turn on, change color, or change intensity of fluorescence in the...Epigenetics with a FRET Based Biosensor Fluorescent Proteins 101: Fluorescent Biosensors ...and the receptor, but readouts could involve fluorescence, pH change, heat, etc. The last component of...readout for whatever the bio-recognition event is (fluorescence, pH, etc.). The transducer will generate a numerical...   Neurotransmitters Many canonical fluorescent as well as FRET-based biosensors have been engineered...the ADP:ATP ratio. Most of these sensors are fluorescence-based and have been optimized for a wide-range...A. V., Shcherbakova D. M., Verkhusha, V. V., Fluorescent Biosensors for Neurotransmission and Neuromodulation...

...observed with fluorescence microscopy. This application of AHA labeling is known as fluorescence noncanonical...localization of protein synthesis. Alternatively, fluorescent protein tags can be used to facilitate the visualization...protein synthesis in situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage...there is the question of whether or not the tags influence behavior. A conference and a breakthrough! During...covalently links the azido containing AHA to a fluorescent alkyne tag. The newly synthesized and tagged ...amplification. The amplified DNA segment is labeled with fluorescent probes that enable visualization of the location...PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which is best for me?...

...Addgene’s Fluorescent Protein Plasmids and Resources Relevant Blog Posts Plasmids 101: Green Fluorescent Protein...localization studies or make a reporter construct. Fluorescent proteins (FPs) are also the foundation of multiple...use in biological research. Photoconvertible fluorescent proteins Newly developed photoconvertible FPs...Sattarzadeh et al., as well as a wide variety of other fluorescent proteins for your imaging needs. If you need ...LL. A bright and photostable photoconvertible fluorescent protein. Nat Methods. 2009 Feb 6(2):131-3.  PubMed...Verkhusha VV. A photoswitchable orange-to-far-red fluorescent protein, PSmOrange. Nat Methods. 2011 Jul 31;...Protein Which Fluorescent Protein Should I Use? ...

...nanobody conjugated with Alexa Fluor 488, Alexa Fluor 568, or Alexa Fluor 647 before application to the...Western blots. ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically... distance between the target antigen and the fluorescent label, which can reduce the potential image resolution... resolution of super-resolution fluorescence microscopy. Their size also limits their ability to penetrate... This process can be avoided direct antibody fluorophore conjugates are used, but these are usually more...penetration and decreases the distance between a fluorescent tag and the target antigen to about 2 nm, which...secondary nanobodies conjugated with the desired fluorophores. In figure 4B below, triple colocalization is...

...the Addgene blog Fluorescent proteins 101: Fluorescent Biosensors Which Fluorescent Microscopy Technique...circularly permuted green fluorescent protein (GFP), and nLightR uses a red fluorescent protein, mApple. Kagiampaki...findings that red fluorescent proteins tend to be less sensitive than green fluorescent proteins. nLightR..., J., & Li, Y. (2019). A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection...Plasmids Find Optogenetics Plasmids Visit the Fluorescent Protein Guide Pages ...

...has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011). This technique is performed...the semipermeable membrane from the extracellular fluid into the perfused liquid. The probe then collects...this, researchers from some labs have developed fluorescent-protein based DA sensors (Figure 2). The dLight1... sensors, the new dLight1 sensor is a single fluorescent protein-based sensor. This sensor was engineered...a GFP variant and a DA receptor such that GFP fluoresces when the receptor is bound by DA. Two variants...affinity and high-magnitude responses (increases in fluorescence) at low concentrations (e.g., 100 nm) of DA....variant bound to a DA receptor, such that the GFP fluoresces upon DA-receptor binding (Sun et al., 2018). ...

...stranded DNA bound by tTALE-FP with the fluorescent protein fluorescing green. Image from Shin et al., 2019...intercalating fluorescent dyes for DNA-protein interaction studies? Kyubong Jo's lab generated fluorescent protein...this vector is suitable for flow cytometry and fluorescence microscopy, they also show that this method ...dx.doi.org/10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining dye Article...specific DNA sequences which brings the attached fluorescent protein to the DNA. They created plasmids for...tTALE-mCherry. In comparison to intercalating fluorescent dyes which can cause photo-induced DNA breakage... lab also used tTALE-FP in a single molecule-fluorescence assay to monitor restriction enzyme digestion...

...work!   Fluorophores When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein...us that, although fluorescent proteins are incredibly useful, chemical fluorophores can still be used ...Better Dyeing Through Chemistry & Small Molecule Fluorophores In this post, Luke Lavis from Janelia Research...variety of experiments. Read more to find chemical fluorophores for your next experiment. Nucleic acid based...to different pH’s and some that are fused to fluorescent proteins. She also showed that R-bodies can rupture...to achieve more efficient genome editing - all influenced by an understanding of Cas9 biophysics! Casilio...genes in a single cell, and localization of a fluorescent signal to your target gene. Read this post to...

... of fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein... Cre-loxP system to express one of the three fluorescent protein (XFP) genes in a transgene. When multiple...Sakaguchi et al., 2018) (Figure 1). Once introduced, fluorescent proteins were best imaged after tissue clearing... expression levels. Clearing large tissues - Fluorescent proteins are very stable in SeeDB2. However, ...matching of tissue samples for High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018)...Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system." Nature 450.7166...multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags." eLife 7 (2018): ...

...peak fluorescence and resting fluorescence (achieved through lowering sensor resting fluorescence, while...scientists access to a wide array of promoters and fluorophores for validating and testing purposes. We will...approximately twice as bright, relative to baseline fluorescence, as the GCaMP7f variant, and is approximately...variant. jGCaMP7b exhibits the brightest resting fluorescence and can be used for imaging small neuronal processes...while maintaining high peak fluorescence) and is useful for imaging signaling activity in large populations...of densely-labeled neurons because background fluorescence from inactive neurons is reduced. This variant...localization should improve the specificity of influence measurements by reducing photostimulation of the...

...apart. While the classic Green Fluorescent Protein (GFP) and other fluorescent proteins revolutionized live-cell... with a fluorescence-based approach. These challenges can include background autofluorescence from the...yields results that don't reflect true biology. Fluorescent fusions utilizing bulky tags can disrupt normal...can suffer from photobleaching and cellular autofluorescence (Tsien, 1998). What sets HiBiT apart from ...including NanoBRET assays to monitor the binding of fluorescently-labeled VEGF-A to the heteromeric receptor complex..., H. (2017). Assessing phototoxicity in live fluorescence imaging. Nature Methods, 14(7), 657–661. https...2021). Use of NanoBiT and NanoBRET to monitor fluorescent VEGF-A binding kinetics to VEGFR2/NRP1 heteromeric...

... curve with known amounts of protein. Fluorophores Fluorophores are light-sensitive molecules which emit...cell or tissue sample. Figure 6: Immunofluorescence, in which fluorophore-conjugated primary antibodies ...label cells in a sample. Immunofluorescence (IF) This method uses fluorophore-conjugated antibodies to...IF uses fluorophore color to differentiate between the targets of interest, if your fluorophore is bound...wells) by fluorescence activated cell sorting (FACS). Figure 7: Flow cytometry uses fluorescently-labeled...the fluorophores used is important. It is best to inversely pair the brightness of the fluorophore with...If a bright fluorophore is on an abundant target, it obscures fainter signals. Fluorophores can emit signals...

...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; ...