We narrowed to 478 results for: FLU

...maintenance phase and do not allow cells to reach 100% confluence (80–90% is ideal). Pass and plate the cells the...from 2 x T-175 flasks. Cells should be at ~80% confluence. For each T-175 flask: Aspirate culture media...resuspend cells from the CS2. Cells should be at ~80% confluence. Aspirate culture media and rinse once with 60...Sample Data Figure 1: HEK293T cells at various confluencies. This AAV production protocol should be started...started with cells that are ~80% confluent (center panel). These cells were imaged with a 10X objective ...

...with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment Equipment...batch of PEI is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the ...efficiencies as can be seen here by the amount of green fluorescent protein expression (green in the right panels...

...the individual wells become confluent, expand into larger vessels. A confluent well of a 6-well dish can...can be expanded into a 10 cm dish. A confluent 10 cm dish can be expanded into two 75 cm 2 flasks, etc....

...the next day they will be approximately 50–60% confluent. For 293T cells this is about 2 × 10 6 cells in...Do not use the medium if the cells are overly confluent or the medium is acidified (i.e., the medium appears...cells have expanded but before they become over-confluent, trypsinize the cells and expand them to larger...

...cells from incubator. The cells should be 50-80% confluent and in DMEM that does not contain antibiotics....The target cells should be approximately 80-90% confluent. c. Dilute puromycin in the preferred culture ...: b. Target cells should be approximately 70% confluent. Change to fresh culture media containing 8 μg...

...transfection protocol Fluorescence Titering Assay for Lentivirus Quantify virus using fluorescence measurements...

...ROX dye), to normalize non-PCR–related fluorescence fluctuations and to minimize well-to-well variability...

...batch of PEI is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the ...

...different concentrations of DNA. In our hands fluorescent dye methods tend to report lower values than ...

... Colony Formation Titration Assay Lentivirus Fluorescence Titration Assay AAV Titration by qPCR Addgene...

...  APCAlexa Fluor 647   Alexa Fluor 700APC-H7APC-Cy7   Many factors can influence the level of... you'll need to pick a fluorophore for each target and ensure the fluorophores, together, are able to ...reporter's fluorescence. Another way to label proteins is through treating cells with fluorophore-conjugated...dead cells. The combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your...outlining a few commonly used fluorophores and their emission spectra. Some fluorophores carry their emission ...and RB545. Fluorophores can also come in combination of two conjugated single fluorophores, like APC-Cy7...respective colors of a few commonly used fluorophores. Each fluorophore has a defined emission maximum. Note...

...fusion protein called CalFlux VNT. CalFlux VNT is composed of Venus fluorescent protein, the Troponin ...NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...better biosensors. NanoLuc®-fluorescent protein fusions for imaging Fluorescent proteins are fantastic imaging...couple Nluc with a red-shifted fluor. Here, NLuc internally excites the fluor through a process known as ...BRET). The fluor then emits light at a tissue-penetrating wavelength above 600 nm. LumiFluors Schaub, F.X...more penetrative orange fluorescent protein fusion (OgNLuc). Both fluorescent proteins can be excited ... the lab has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed...

...the associated fluorophore or stained with the associated fluorochrome, emits a fluorescent signal that ...the level of fluorescence. A cell expressing high levels of the fluorophore or fluorochrome-stained protein...antibody conjugated fluorophores Fluorescent antibodies get around the challenges of fluorescent protein fusions...such as fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy... have a variety of fluorescent light sources. When interrogated with a fluorescent light source, a cell... to a fluorochrome. In indirect staining a primary antibody binds to a target and a fluorochrome-conjugated... panel of “fluorescence-minus one” controls, samples stained with all of the fluorochromes in the experiment...

...non-radiatively from an excited molecular fluorophore (the donor) to another fluorophore (the acceptor) by means of ...the two fluorophores used are in close vicinity. Thus, the distance between the two fluorophores and their... of a fluorophore pair, donor:acceptor stoichiometry, and cross-talk between the two fluorophore colours... filters chosen for the donor fluorescence and the acceptor fluorescence. Additionally, this method could... the fluorescence decay time of the donor. When FRET occurs between the pairs, donor fluorescence is quenched...factors, such as pH or autofluorescence background, can change the fluorescence decay time and have to...course talking about the now well-known FRET, aka Fluorescence (Förster) Resonance Energy Transfer, technique...

...for autofluorescence extraction Cells produce a low level of fluorescence called autofluorescence (you...to identify a fluorophore, spectral flow cytometers use all detectors for all fluorophores. This allows...tool called “autofluorescence extraction” which allows the user to use the autofluorescence in a sample...sample as a fluorescent marker. Given that the level of autofluorescence across various tissue types varies...you wish to use the autofluorescence extraction feature. Although autofluorescence extraction is an optional...samples stained with a single fluorophore. They’re required for every fluorophore you use in your panel, whether... the spectral signature of a fluorophore itself from the autofluorescence of the beads or cells. Overall...

... blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein...area with lots of bright new plasmids is our Fluorescent Biosensors Collection — scientists are always...measure! Let’s take a look at some of the hottest fluorescent biosensors new to Addgene this year. Feel the...Hiroshi Hibino and colleagues devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular...region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent protein is inserted into one of the loop regions...  Find MTRIA sensors here!   Ino D, et al. A fluorescent sensor for real-time measurement of extracellular...co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma membrane only when the...

...More about Fluorescent Protein-Based Tools Learn Some of the Pitfalls When Using Fluorescent Proteins Control...Resources on Asddgene.org Find Fluorescent Protein Plasmids Find Fluorescent Protein Kits Find Empty Backbones...historically been difficult, as techniques like fluorescent in situ hybridization (FISH) and chromosome capture...be used in vivo. Some researchers have used fluorescently tagged DNA-binding proteins to label certain...chromosome. This image represents the composite of fluorescent measurements from red, blue, and green channels...targeting capacity. By using a gRNA to target fluorescently labeled dCas9 to a given genomic locus, you ... thereby decreasing the amount of background fluorescence from unbound GFP-dCas9. With these improvements...

...than any fluorescent protein earlier described. “Until our proteins, there were lots of fluorescent proteins... If you start poking around on Addgene’s Fluorescent Protein Guide to In Vivo Imaging, you’ll pretty ...good reason. We all know scientists have used fluorescent proteins to observe what’s happening inside cells... of decades. Green is the classic color, but fluorescent proteins are available in a variety of hues. ...unimpeded. The challenge then was to develop fluorescent proteins that would fall within that near-infrared... from jellyfishes and corals, but all of them fluoresce outside the transparency window of mammalian tissues... Verkhusha said. “We developed near-infrared fluorescent proteins within this transparency window, so ...

...years — a green fluorescent protein, a red fluorescent protein, a fancy new green fluorescent protein, some...new-and-improved fluorescent protein you want to try, just swap in a new fluorescent ligand and see the...purification tags, another version with that same red fluorescent protein somebody made because they forgot it ...of chemical dyes provide many advantages over fluorescent proteins, opening up new applications.   ...strategies. The green structure represents a fluorophore or other useful ligand, gray structure is the...biarsenical dyes such as FlAsH (green, based on fluorescein) or ReAsH (red, based on resorufin). SNAP-tag...dihydrofolate reductase (eDHFR) could be labeled with fluorescent derivatives of its substrate trimpethoprim (TMP-tag...

... conjugated to a fluorophore, not every marker will have a corresponding fluorophore-conjugated antibody...cases, fluorophore-conjugated antibodies may exist, but only with a limited selection of fluorophores. If... tag but not a fluorophore. In a separate step, cells are incubated with a fluorophore-conjugated streptavidin...wide range of fluorophores, allowing for great flexibility when it comes to fluorophore selection for ...common method is a cell surface label, where fluorophore-conjugated antibodies directly bind to epitopes...employ beyond simply incubating cells with single-fluorophore-conjugated antibodies. These methods can include...interaction (Figure 1a). This interaction brings the fluorophore in proximity to the target marker, allowing for...

...overcome the issues with fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash...from the fluorophore is pH-dependent. Signal is simultaneously acquired from the the fluorescent and luminescent...mimetic and induces fluorescence in the presence of DHFBI, a small molecule fluorophore. When fused to an...FRB-NES and Cas9(C)-FKBP-2xNLS dimerization and net influx into the nucleus, where the functional Cas9 molecule...visualization of single molecules via targeted fluorescent molecules. The Vale lab and the Weissman lab ...protein of interest are recognized and thus fluorescently labeled by scFv antibodies fused to sfGFP. This... This system amplifies the intensity of fluorescence signal and enables tracking of single molecules within...

...challenging due to low fluorescence intensity, caused partially by fluorophore photoinstability, as well... by immunostaining. Although the fluorophores used are fluorescently distinct, their protein sequences...cells. The Brainbow-1.0 construct contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (...recombination. Brainbow-1.1 scales up the system to four fluorescent proteins, with recombination events specified...but the principles remain the same. The first fluorescent protein in the construct, OFP (orange), is expressed...two tandem invertible segments, each with two fluorescent protein coding sequences in opposite orientations...expressed; once Cre is removed from the system, the fluorescent protein whose coding sequence is closest to the...

...like chromoproteins, fluorescent proteins, and luminescent proteins. While fluorescent proteins and luminescence...from fluorescent proteins? Before going into the difference between chromoproteins and fluorescent proteins...the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to blue ...light, whereas chromoproteins do not fluoresce at all or fluoresce at very low levels. Chromoproteins in...detection. Unlike fluorescence or luminescence, which require UV lamps, fluorometers, or luminometers,...The ocean is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful... chromoproteins are actually a subset of the fluorescent protein family. What sets them apart is the way...

...with rhodopsin quenching the added fluor for this two-part fluorescent system, and rhodopsin still functioning... elements: a voltage-sensing component and a fluorescent output. The GEVI family of gadgets includes Ace...Chemigenetic sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing...VSD-based sensors consist of a VSD fused to a single fluorescent protein (FP). This class of GEVI was the earliest...of red-shifted indicators, and brightness of fluorescence all have room for improvement. These sensors...rhodopsin functions as both the voltage detector and fluorescent signaler. The pro of this system is the kinetic...were initially evolved for ion transport, not fluorescence, they are not nearly as bright as the other ...

...a magnetoresponsive fluorescent protein. Controlling an optogenetic or fluorescent tool with magnets would... do fluorescent proteins respond to magnetic fields?” and “What are magnetoresponsive fluorescent proteins...set out to do just that — by developing a new fluorescent protein tool that responds strongly to magnetic...Motivated by curiosity Although a wide range of fluorescent proteins and optogenetic tools exist, humans ...responses of cryptochrome proteins or organic fluorophores (Karki et al., 2021; Lee et al., 2011; Matsuoka...EGFP — showed a slight but detectable change in fluorescence when a small magnet was passed over the sample...sample (Frank Hayward et al., 2024). The fluorescence intensity decreased whenever the magnet approached...

... Since fluorescent proteins (FP) require external light activation, you can’t use fluorescence to monitor...detection is difficult using fluorescent imaging due to autofluorescence and a high signal-to-noise ratio... Learn about Photoactivatable Fluorescent Proteins Check Out Fluorescent Protein Timers Use Rosella to...All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein...Fluorescent imaging techniques have become indispensable tools for molecular and cell biologists over...experimental success can be affected by both autofluorescence and photobleaching. Researchers have long ...luciferase transfers energy to the FP, resulting in a fluorescent signal. Since their first publication in 2012...

...intrinsically fluorescent and can be detected with highly epitope-specific, fluorescently-labeled antibodies...usually provided by red fluorescent proteins such as tdTomato or RFP or blue fluorescent proteins like Cyan... and David W. Piston. "Fluorescent protein tracking and detection: fluorescent protein structure and color...tracers Neuronal tracers such as epitope tags and fluorescent proteins (FP) have also been important in mapping...tag-specific antibodies. The most famous ones are influenza hemagglutinin (HA), myelocytomatosis viral oncogene...compared to epitopes tags, FPs are intrinsically fluorescent and can therefore be used in live imaging experiments...tracer With the limitations of epitope tags and fluorescent proteins, it has been crucial to develop new ...

...up new fluorescent markers before deciding to use them, using a tool like BioLegend’s Fluorophore Guide...the process of correcting spillover from one fluorescent channel to another. When you label your samples...samples with multiple antibodies, the fluorescent probes on the antibodies may have similar emission spectra...spectra, meaning they will emit fluorescent light at similar wavelengths (See this handy emission spectra...spectra). Your cytometer will therefore record the fluorescence for both at similar levels, meaning that your...desired cell population. By compensating between fluorescent channels, you can correct for this spillover ...your flow panel, you will need to know both the fluorescent markers you intend to use and your cytometer’...

...inhibitory synaptic proteins Fluorescent protein system for C. elegans Fluorescent timers for studying cell...Split-wrmScarlet, a fluorescent proteins system for Caenorhabditis elegans By Jennifer Tsang Fluorescent proteins...are not fluorescent on their own. When the two fragments come together, they fuse and fluoresce. The team...neurotransmitters. Using genetically encoded fluorescent proteins coupled with Fibronectin intrabodies...localize to the synapse and minimize extraneous fluorescence from other regions of the cell. By combining...combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory synapses or that...Predicting cell cycle length with a histone-fused fluorescent timer By Kate Harten DeMaio Studies of the cell...

...cleaves the supplied fluorescently labelled ssDNA substrates. Upon cleavage, the fluorescence is no longer quenched... cleavage of fluorescently labelled RNA, the authors employ a P51 cardboard fluorescence visualizer that...streptavidin portion of the probe via biotin. The fluorescent portion of the probe that is cut will continue...cleaves a nearby RNA sequence that contains a fluorophore that is quenched when intact and released when...when cleaved. Cleavage results in fluorescence and this approach can be incorporated into a lateral-flow...recombinase, and the ssDNA-FQ reporters that exhibit fluorescence upon cleavage. Upon amplification by the RPA...cleavage of the ssDNA-FQ reporter resulting in fluorescence. Cas12a binds all the amplified target regions...