We narrowed to 478 results for: FLU

... Protocols Fluorescence Titering Assay Fluorescence Titering Assay ...resulting in 25% fluorescent cells, then the titer is: (150,000 cells x 0.25 fluorescent x 100)/(1.5 mL)...resulted in 25% fluorescent cells, then the titer is: (150,000 cells x 0.25 fluorescent)/(0.015 mL) = ...preparations carrying a fluorescence marker. Protocols...to titer lentiviral preparations that carry a fluorescence marker. Determining the titer of your lentiviral...uses microscopy to determine the percentage of fluorescent cells, these cells can still be used in downstream...media, replace with fresh media Day 4–5: Count fluorescent cells Equipment Class II, Type A2 Biological ...

...RFP fluorescence suggests an increase in autophagic flux, while an increase in GFP fluorescence relative...of several fluorescent autophagy reporters now allows for the measurement of autophagic flux, or the changes... to emission of only red fluorescence. The ratio of red to green fluorescence allows for clear distinction...control. Autophagic flux is estimated by the GFP/RFP ratio: a decrease in GFP fluorescence relative to RFP...relative to RFP fluorescence suggests a decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at...photoconversion Light sheet fluorescence microscopy Resources on Addgene.org Fluorescent protein plasmids and...autophagy biosensor that’s comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant...

... of fluorescently-labeled components Fluorescence-activated cell sorting (for cells); fluorescent HPLC...and fluorescent reporters being especially popular. We’ll give you an introduction to fluorescent biosensors...calcium and zinc. Some biosensors contain a single fluorescent protein fused to a domain that senses the concentration...When calcium is absent, the construct does not fluoresce; when present, calcium induces CaM/RS20 interaction...s structural organization and activating the fluorescent signal. Blue flame GECO biosensors from Robert...s lab display calcium-dependent increases in fluorescence of up to 11,000-fold. FRET biosensors are another...sensors created by Roger Tsien’s lab contain two fluorescent proteins (a FRET pair) separated by CaM and M13...

... as fluorophores in several different ways. Researchers have used oligonucleotides in fluorescent visualization...decades, most notably for fluorescent in situ hybridization (FISH) where fluorophore-linked antibodies are...applications including aptamer based fluorescent biosensors. Aptabeacons Fluorescence resonance energy transfer ...state contains a fluorophore and a quencher in close proximity. For instance, the fluorophore and quencher...aptamers fluoresce until they bind to the appropriate ligand, but relying on a lack of fluorescence for readout...applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing with veggies) Though the...is bound to a protein fluor. Transcribing and translating a protein fluorophore also takes time - bypassing...

..., like luciferases, as well as fluorescent ones, like Green Fluorescent Protein (GFP). These molecules...previously reach. Nobel Prize for fluorescent proteins The importance of fluorescent proteins was recognized in...learning more about fluorescent proteins, I invite you to download our Fluorescent Protein eBook and learn...laboratory. The discovery of GFP The story of how fluorescent proteins like GFP came to be used as research...function and shift its emission spectrum. New fluorescent proteins with new colors Around the same time...hunch paid off, and he was able to report new fluorophores like DsRed from Discosoma sp. and Katushka/mKate...quadricolor. With his discoveries, another part of the fluorescent protein puzzle fell into place, as they, taken...

...time.Hallmarks of fluorescent protein timers When FP timers are first expressed, they predominantly fluoresce one ...independently) originally fluoresce green or blue and, when they mature, fluoresce red. The ratio of the ...Advice on Choosing Your Fluorescent Protein Resources at Addgene.org Find Fluorescent Protein Timers Browse...Browse All Fluorescent Protein Empty Backbones Check Out Our Fluorescent Protein Guide ...Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell...situ hybridization experiments. In many cases fluorescent proteins have usurped old methods or complemented...color but slowly mature until they predominantly fluoresce a second color. This change is usually hypothesized...

...from low fluorescent states to high fluorescence states, they can change from one fluorescent color to...proteins. Photoactivatable fluorescent proteins (PA-FPs) are fluorescent proteins that display unique...their fluorescence can be switched on and off reversibly. This ability to modulate fluorescence allows...switched on from a state of dim or no fluorescence, to a brighter fluorescent state by exposure to a specific...photoconvertible fluorescent proteins (PC-FPs). PC-FPs can convert from one fluorescent color to another... a bright fluorescent state (“kindling”), and from a bright state to a dark non-fluorescent state (“quenching...reversible, monomeric red fluorescent PA-FPs. rsCherries have a high background fluorescence, but their single...

...Bioluminescence and fluorescence from proteins such as Green Fluorescent Protein (GFP) has likely existed...iterations of fluorophores are constantly being engineered. Table 1 lists a few common fluorescent proteins...specific mutations comprising common fluorophores Fluorescent Protein Mutations Relative to Wildtype...folding, maturation, and fluorescence. Biosensors: A wide array of GFP-based fluorescent biosensors has been...techniques in fluorescent microscopy. Read More: All Plasmids 101 Articles Addgene's Fluorescent Protein ... its biochemical properties. Now GFP and its fluorescent derivatives are a staple in the lab. GFP is used... common applications in the lab. Why Green Fluorescent Protein? GFP is a ~27 kDa protein consisting ...

...opportunities. Xin Zhou: Fluorescent Light-inducible Proteins (FLiPs) Fluorescent proteins can be applied...Brightest Fluorescent Protein for Your Experiments Resources on Addgene.org Find Fluorescent Protein ...Fluorescent proteins have enabled scientists to pursue creative research avenues previously unavailable...applications, researchers are finding new ways to apply fluorescent proteins everyday.  The late Michael Davidson... and develop these tools by making their own fluorescent protein technologies widely available to the ... are intended to help scientists share their fluorescent protein research with the academic community ...stood out for their creativity in the use of fluorescent proteins and the potential for their work to ...

...track fluorescently labeled cells? 1. Epifluorescence microscopy This technique uses a fluorescence microscope...for many fluorescent proteins. Additional fluorescent protein tools Photoactivable fluorescent proteins...the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter... By using cells labelled by fluorescent chemicals or by other fluorescent proteins, labs were also able... that exist in vivo. The use of fluorescent pathogens and fluorescent cells enables scientists to monitor...Laser include: Blue-green fluorescent proteins like EGFP Yellow-orange fluorescent proteins like TagRFP, ...irreversible photoconversion from green fluorescence to red fluorescence) are also useful tools to use with...

...turn up the brightness in your fluorescent imaging experiments. Fluorescent protein fusions One of the easiest...Blog Sign Up to Receive the Fluorescent Protein eBook Avoid Fluorescent Protein Oligomerization Learn...Learn about Small Molecule Fluorphores Resources on Addgene.org Find Fluorescent Protein Empy Backbones ...biological lesson and created a system to amplify fluorescent signals. Named for the "stellar explosion SUperNova... to track a given protein is to fuse it to a fluorescent protein. You can then study where the protein...these problems? Instead of directly fusing a fluorescent protein to your protein of interest, you instead.... This system amplifies the intensity of the fluorescent signal and enables single molecule tracking  ...

... using fluorescence (for some background on fluorescence, try the Introduction to Fluorescence Microscopy...proteins. You can then use fluorescent microscopy to look for those fluorescent molecules, knowing that ...to a fluorophore (Figure 1).   Figure 1: Comparison between how direct immunofluorescence and indirect...Introduction to Fluorescence Microscopy. iBiology. https://www.ibiology.org/talks/fluorescence-microscopy/...Antibodies Learn more about fluorescence microscopy in Which Fluorescence Microscopy Techniques is Best...Immunofluorescence (IF), is an immunoassay that brings to light the cellular world. The technique allows...and performing an IF experiment. What is immunofluorescence? Generally speaking, IF is an application...

...Properties of luminescence versus fluorescence   Luminescence Fluorescence Source of light Chemical reaction...of energy from an excited donor fluorophore to an acceptor fluorophore, which then emits it as light. ...brings the fluorescent protein close enough, energy is transferred by BRET and the fluorescent protein emits...common genetic reporters and alternatives to fluorescent proteins. Figure 1: Firefly, from Wikimedia...the title of this post, this reaction is not fluorescence! Chemiluminescence, which includes all bioluminescence...of light by a chemical reaction. In contrast, fluorescence occurs when a molecule is excited by a high-...to be provided and are generally dimmer than fluorescent proteins. On the other hand, they don’t require...

...reagents like fluorescently-labeled nanobodies or scFvs. Many, many options for fluorescently conjugated ...then label that with fluorescently-conjugated streptavidin to get more fluorophores per antibody, or conjugate...Antibodies Which Fluorescence Microscopy Technique is Best for Me? Introduction to Immunofluorescence Resources...You may be familiar with immunofluorescence (IF, often referred to as immunocytochemistry (ICC) when ...binds a target protein in your sample, then a fluorescently labeled secondary antibody binds that first ...The general process is similar to standard immunofluorescence, but with some extra considerations (Im et...  Figure 1: Schematic of multiplex immunofluorescence approaches. A) Sequential Multiplex IF, where...

...the fluorescence of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified...small molecule activates the fluorescence of the small molecule. Fluorescent RNA aptamers are used to track... applications of fluorescent RNA aptamers in Addgene’s collection. Green fluorescent aptamers Spinach ...selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution...blog Learn more about aptamer fluorophores Investigate other fluorescent biosensors Find other uses for...using aptamers might know this best, as most fluorescent aptamers are named for a fruit or vegetable! ...What is an RNA aptamer? RNA is not inherently fluorescent and can be challenging to track intracellularly...

...Developments in light sheet fluorescence microscopy Modern light sheet fluorescence microscopy was first pioneered... advances in microscopy has been light sheet fluorescence microscopy (LSFM). The underlying method was...colleagues and originally named orthogonal-plane fluorescence optical sectioning (OPFOS) (2). Arne Voie, David...laser beam into a thin sheet to illuminate a fluorescent sample and captured the reflected light using...configurations’ various features see (4,7).   Light sheet fluorescence microscopy vs. confocal microscopy In confocal...However, the excitation light excites all of the fluorophores as it passes through the specimen, which often...obstacles (e.g. air bubbles or high concentration of fluorophores) cast shadows along the illumination path. The...

... months ago, we shared an introduction to immunofluorescence (IF) - a common method for visualizing molecules...1985) Postfixation detergent treatment for immunofluorescence suppresses localization of some integral ...Yong WH (2018) An Introduction to Performing Immunofluorescence Staining. Methods in Molecular Biology. Springer...Addgene blog: Antibodies 101: Introduction to Immunofluorescence Antibodies 101: Epitope Availability Antibodies...

...in a change in the fluorescent readout. In this blog we will focus on two fluorescent CRISPR reporter methods...surpass GEmCherry2 in background fluorescence and in-frame fluorescence.  Figure 2: The GEmCherry...assays. Recently scientists have created CRISPR fluorescent reporter assays to quantify targeted Cas9 cleavage...enrich for mutants through the presence of a fluorescent reporter. Lastly, once a mutant is identified...at Addgene GEMCherry2, an improved monomeric fluorescent protein for assessing CRISPR/Cas9 activity Guide...but most are labor-intensive, require multiple fluorescent proteins, and aren’t that efficient. To overcome...Denham lab developed a modified monomeric red fluorescent protein (RFP), named GEMCherry2, that can rapidly...

...emission by fluorescence. The widespread use of FRET with fluorescent molecules, including fluorescent proteins...donor fluorophore to the acceptor fluorophore. The typical steps in FRET are: 1. Donor fluorophore excitation...acceptor fluorophore--think of it as a virtual photon 3. Relaxation of the acceptor fluorophore back to...acceptor fluorophores must be compatible, such that the emission spectrum of the donor fluorophore overlaps..., et al. "Fluorescent protein based FRET pairs with improved dynamic range for fluorescence lifetime measurements...Miyawaki, Atsushi, et al. "Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin... Out Fluorescent Protein Biosensors Browse the Entire Fluorescent Protein Collection ...

... is the permanent fluorescent tag portion of Rosella: it will emit red fluorescence regardless of its ...the fluorescent properties of Rosella.   Table 2: Effect of Localization on Rosella’s fluorescence emission...green fluorescence accumulates in the vacuole. See Figure 2 for an example of what fluorescence looks ...Photoconversion Light Sheet Fluorescence Microscopy Additional Resources on Addgene.org Fluorescent Protein Plasmids...Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, another fluorescent pH-biosensor for studying...Rosella is a pH-sensitive fluorescent biosensor that was recently deposited with Addgene by Dr. Mark ...Rosella’s autophagy-sensing abilities is that its fluorescence emission spectra changes when it goes from a...

...containing several types of fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP) tags, a selectable...Blog Learn about Fluorescent Protein Timers Check out Photoactivatable Fluorescent Proteins Use Rosella...SapTrap Kit Page Find Other Fluorescent Protein Kits Browse the Fluorescent Protein Collection ...GFP over 50 years ago, the growing spectrum of fluorescent proteins (FPs) has been an invaluable resource...limitations to working with FPs. Overexpression of fluorescently tagged proteins can lead to improper protein...One way to avoid the pitfalls of overexpressed fluorescent protein fusions is to replace the genomic copy...copy of your gene of interest with a fluorescent protein fusion. CRISPR genome editing can help accomplish...

...super-resolution (3). Fluorescence microscopy techniques   Fluorescent Widefield Microscopy Fluorescent Widefield... unique characteristics of fluorophores to block out of focus fluorescence from reaching a detector or...Internal Reflection Fluorescence (TIRF) microscope can be used. TIRF only excites fluorophores within a few ...Choosing Your Fluorescent Proteins for Multi-Color Imaging Choose the Brightest Fluorescent Protein for ... Addgene.org Browse Our Fluorescent Protein Resources Find Empty Fluorescent Protein Backbones Find Plasmids... into noise. Due to its unrivaled contrast, fluorescence imaging has emerged as the dominant light microscopy...modern biology (1). When performed correctly, fluorescence microscopy provides a high contrast image in...

...Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how fluorescent proteins glow?   ...the protein is non-fluorescent, thus allowing very low background level of fluorescence. Examples for such...Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing a fluorescent protein...you, "Which fluorescent protein should I use?" And if you're looking for more fluorescence microscopy ...Interactive Visualization of Fluorescent Protein Properties  Fluorescence Spectrum Viewer  from BD bioscience...   Fluorescent Proteins (FPs) were first discovered over 50 years ago, with the discovery of the Green...Green Fluorescent Protein (GFP), a protein from the jellyfish Aequorea Victoria. Since that discovery, the...

...College of Medicine. Several sets of near-infrared fluorescent proteins (NIR FPs) and biosensors have been created...Addgene. Biliverdin chromophore Near-infrared fluorescent proteins (NIR FPs) were derived from natural ...their molecular brightness (a product of the fluorescence quantum yield and the extinction coefficient... FPs are useful in vivo probes, due to low autofluorescence, low scattering, and deep penetration of red-shifted...reporters can tell us about cellular events via  fluorescence changes occurring due to protein synthesis, ...mammalian cells. dDetermined as effective NIR fluorescence in transiently transfected live HeLa cells with...supply of exogenous BV and after normalization to fluorescence of co-transfected EGFP. e Originally reported...

...photostable fluorescent protein. Photobleaching is the irreversible destruction of a fluorophore under the...the influence of light. Any fluorescent molecule will photobleach at some point. For live-cell imaging...desirable to have fluorescent proteins that are photostable. On top of photobleaching, fluorescent proteins may... the ideal situation, a fluorescent proteis should emit a stable fluorescence signal, showing no or little...power is a property of the fluorescent protein. This can differ between fluorescent proteins in the same spectral...performed that measures fluorescence intensity over time. In order to predict how fluorophores perform in ‘real...measurements for different fluorescent proteins and by comparing how the fluorescence intensity changes over...

...biophysical interaction between two fluorophores. When the first fluorophore (the donor) is excited, it can...acids/lipids of interest are fluorophore labeled and that these fluorophores can undergo FRET when they ...targeting a FRET fluorophore to a biomolecule is a bit like a Fedex delivery. Your fluorophore package needs...there is no fluorescence background resulting from nonspecific donor labeling. Fluorescent proteins (FPs...incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (...Experiments Beware of Fluorescent Protein Aggregation Brush up on Your Fluorescent Protein Basics Resources...Browse Empty Plasmids to Fluorescent Protein Fusions Check out Our Fluorescent Protein Guide ...

...assemble the full structure of the fluorescent protein, producing fluorescent signal. Image modified from Feng...Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis...use with green-yellow or orange-red fluorogen A split fluorescent reporter with rapid and reversible complementation... protein of interest to each part of a split fluorescent protein (FP) and measure the signal produced ...This method is generally known as Bimolecular Fluorescence Complementation (BiFC). It can provide a qualitative...plasmid for your experiment.  Highlighted Split-Fluorescent Protein Tags Browse the table below, and click...

...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes,...stability of fluorescence over time is crucial. For instance, the fluorescence of fluorescein isothiocyanate...the b(right)est fluorescent protein Fluorescent Proteins 101: A History of Fluorescent Proteins ...dyes, and fluorescent ligands. Figure created with BioRender.com. Fluorescence refers to the emission...spectrum and emission spectrum. Fluorescent dyes utilize these fluorophores by combining them with a molecular...interest. Fluorescence microscopy takes advantage of the characteristics of these fluorophores to enable...can utilize these dyes. The most-used fluorescent dye is fluorescein isothiocyanate (FITC), with excitation...

...position and fluoresce. FlipCherry: Red fluorescence protease reporter To add to the fluorogenic reporter...caspase results in a change in fluorescence emission. FRET and fluorescence based reporters however have...necessary for fluorescence. When these parts are separated from each other, the protein cannot fluoresce. When...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...vivo, scientists have used genetically encoded fluorescent executioner caspase reporters. Several apoptosis...difficult to detect, and thus quantify, due to small fluorescence changes of the donor and acceptor after proteolytic...

...deposit plasmids containing the fluorescent protein mNeonGreen. This fluorescent protein joins mTFP1 and mWasabi...yellow-green fluorescent protein mNeonGreen is a yellow-green fluorescent protein that was derived from ...yellow fluorescent protein wavelengths, it can be imaged either with standard green fluorescent protein... other fluorescent proteins. Find mNeonGreen plasmids here! mTFP1: a monomeric teal fluorescent protein...proteins is that they are monomeric fluorescent proteins. Fluorescent proteins that oligomerize often affect...Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging." ...blog Read our fluorescent protein blog posts Find blog posts about choosing fluorescent proteins for your...