We narrowed to 187 results for: Green fluorescence protein
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TypeCollection...expressed in the retina). Fluorescent Cre: The fusion of Cre to a fluorescent reporter enables visualization...pCMV-cre-K Cre-K CMV Mammalian Green 8395 p210 pCMV-CREM CREM CMV Mammalian Green 8401 p224 pCMV-RFP/CREM floxed...within CREM CMV Mammalian Green 8403 p153 pPr-cre Cre probasin Mammalian Green 8404 p236 pPr-CREM CREM ...probasin Mammalian Green 8405 p240 pPr-CREM/Ins CREM with insulators probasin Mammalian Green 8406 p252 pPr-CREM...Mammalian Green 12494 p260 pCMV-CRE-M-AR(wt) CREM fused to AR ligand binding domain CMV Mammalian Green 12496... a stochastic choice of expression among fluorescent proteins, resulting in the indelible marking of mouse...specific system for controlling gene expression. The protein Cre recombinase recognizes 34 bp loxP sites, and...
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Antibodies 101: Designing Your First Flow Panel
TypeBlog Post...inside of your cells. While proteins themselves can emit intrinsic fluorescence when excited with ultraviolet...amino acids found in all proteins, so you can't distinguish the different proteins from each other. To distinguish... A common method of labeling proteins is to fuse them with a fluorescent reporter, such as GFP or mCherry...through detecting the reporter's fluorescence. Another way to label proteins is through treating cells with...distinguish different target proteins in your flow analysis, you'll need to use fluorophores that attach...mCherry, through genetic modification. As your protein of interest will only occur in combination with that...cells. The combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow... -
Fluorescent Protein Guide: FRET
TypeCollection...Collections Fluorescent Proteins FRET Fluorescent Protein Guide: FRET More Fluorescent Protein Resources...for creating individual fluorescently tagged proteins to study protein-protein interactions and a series...not required to be fluorescent. A variety of organic dyes and fluorescent proteins have been developed...is often used to study (1) protein-protein interactions where each protein is separately fused to a donor...Encoding Fluorescent Proteins for FRET The following plasmids can be used to create a fluorescent fusion...fusion protein with your gene of interest and the listed fluorescent protein. Plasmid Color Expression Description...changes within a protein where the donor and acceptor are both fused to the same protein (also referred ... -
Luminescent Imaging with Nano-lanterns
TypeBlog Post...Learn about Photoactivatable Fluorescent Proteins Check Out Fluorescent Protein Timers Use Rosella to Study...Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein Tags... Since fluorescent proteins (FP) require external light activation, you can’t use fluorescence to monitor...generates much less signal than comparable fluorescent proteins. This weak emission hasn’t stopped some ...luciferase variant are used to excite the fused fluorescent protein. This technique is very similar to the widely...of low-copy number proteins, whereas such detection is difficult using fluorescent imaging due to autofluorescence... fluorescent proteins... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post...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 ...targeting a fluorescent donor to your protein is to hardwire the molecule directly into the protein’s peptide...incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (...during protein translation (Ermolenko et al., 2007). All that is required is that the proteins/nucleic...synthesized target protein molecules contain the donor and, more importantly there is no fluorescence background...resulting from nonspecific donor labeling. Fluorescent proteins (FPs) are the gold standard for genetically-encoded... -
FlipGFP, a novel fluorescence protease reporter to study apoptosis
TypeBlog Post...Addgene blog Read our fluorescent proteins blog posts Download the fluorescent proteins 101 eBook Read our... Browse the fluorescent protein collection Find empty backbones for fluorescent protein fusions Browse...necessary for fluorescence. When these parts are separated from each other, the protein cannot fluoresce. When...caspase results in a change in fluorescence emission. FRET and fluorescence based reporters however have...normal antiparallel position and fluoresce. FlipCherry: Red fluorescence protease reporter To add to the... cleave proteins. This protease reporter based on sfCherry however did not initially fluoresce in HEK293... used in FlipGFP can be applied to other fluorescent proteins to create an array of multicolor reporters... -
Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative
TypeBlog Post... 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...DNA. TALE proteins can bind specific DNA sequences which brings the attached fluorescent protein to the ...10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining dye Article contributed...contributed by Leah Schwiesow Calcineurin is a protein phosphatase that responds to rising intracellular... in the cell, calcineurin dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent...this vector is suitable for flow cytometry and fluorescence microscopy, they also show that this method ... -
Tag Your Favorite Yeast Genes with Ease
TypeBlog Post...direct fusion of fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes ... localization of individual proteins, but also investigate protein-protein interactions via FRET. Sidae... especially useful in protein complex studies where tagging of multiple protein products is desired, as... unique protein fusions. This collection is ideal for tandem affinity purification of protein complexes...advantage of this natural process to integrate protein tags into the genomes of S. cerevisiae and S. pombe.... A given insert will typically contain both a protein modification sequence and a selectable gene product...several ready-to-use, modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection... -
Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes
TypeBlog Post...vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent protein biosensors ...tools The luciferase reporter gene The green fluorescent protein reporter gene Resources on Addgene.org... 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...ARGs) are engineered gene clusters that encode proteins that form bacterial gas vesicles. These ~200nm...200nm long vesicles have hollow interiors and a protein shell that excludes water but is gas permeable. These... -
Plasmids 101: Gene, enhancer, and promoter trapping
TypeBlog Post...way. For example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed...construct can be something phenotypic (such as a fluorescent protein or something species-specific) or a selectable... -
Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles
TypeBlog Post... of the many Fluorescent Proteins (FP) available for microscopy, by cloning your protein of interest into...into a vector encoding a fluorescent tag. Once expressed as a fluorescent protein fusion, it is possible...wild-type protein visualized with a fluorescently-tagged antibody. By employing “marker proteins” which ...your fluoresent protein fused to a signal sequence that traffics the fluorescent protein to your organelle...the function of a protein or its role in a disease, researchers often isolate proteins of interest and ...and transport of the protein within a living cell. Analyzing differences in protein localization and transport...mechanisms and protein function. Getting ready to determine the subcellular localization of your protein of interest... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog Post... constitutively express a red, green, or blue (RGB) fluorescent protein (4a-c). When high doses of this...are easy to trace and not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color...trace individual cell’s axons, particularly the two green neurons near the arrows. When lower doses of virus...likely to be transduced by only one RGB AAV and red, green, or blue are the predominate colors expressed (figure...A cocktail of three AAV vectors expressing RGB proteins under the control of an inducer, i.e. tet-off ...morphology studies when used as part of a two part fluorescent tag-then-express system that allows for controlled... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog Post...b(right)est fluorescent proteinFluorescent Proteins 101: A History of Fluorescent Proteins ...stability of fluorescence over time is crucial. For instance, the fluorescence of fluorescein isothiocyanate...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes,...dyes, and fluorescent ligands. Figure created with biorender.com. Fluorescence refers to the emission...attached to self-labeling proteins. Some common examples of these self-labeling proteins would be SNAP-tag® ...through fluorescent labeling. There are several advantages to using these self-labeling proteins over traditional... traditional fluorescent protein tags, such as higher specificity, increased versatility in probe attachment... -
10 Great Guest Posts We're Thankful For
TypeBlog Post...Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization...Research Campus reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores...different pH’s and some that are fused to fluorescent proteins. She also showed that R-bodies can rupture...Zika virus and, upon binding, produce reporter proteins. These toehold switches can be used to quickly... Read this post from Keith Pardee and Alexander Green (both formerly at the Wyss Institute) to learn how...University, describes R-bodies, “force generating protein machines that can extend and retract over many ...Casilio, a system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety... -
Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools
TypeBlog Post...TagRFP-T fluorescence intensity due to interactions with photoactivatable fluorescent protein Dronpa. ...FLIM-FRET acceptors, ShadowG, a novel dark green fluorescent protein (GFP) with an absorption peak of 486 nm... dark yellow fluorescent protein (YFP) with an excitation peak of 519 nm. These proteins have improved...basic-helix-loop-helix) protein. After fusing the CRY2 and CIB1 domains to separate proteins of interest, a researcher... oligomerization when fused to a protein of interest. Fusion protein oligomerization happens within seconds... in TagRFP-T fluorescence intensity at different time points, the authors tracked protein activity while...phosphatase activity, second messenger activity, and protein-protein interactions. We’re excited to see how you ... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post... the Brightest Fluorescent Protein Resources on Addgene.org Find Fluorescent Protein Plasmids Check ...components: a GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP...NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP...which contains the tethering sites for the fluorescent proteins and is driven by an inducible promoter (... PP7 and MS2 coat protein binding sites to tether many copies of the FP-coat protein fusions to their ...RFP-labeled osk and Oskar protein were detected at the posterior pole. Oskar protein levels were negatively... to monitor local translation and determine if proteins are targeted to their destination co- or post-... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog Post...inside of your cells. While proteins themselves can emit intrinsic fluorescence when excited with ultraviolet...amino acids found in all proteins, so you can't distinguish the different proteins from each other. To distinguish... A common method of labeling proteins is to fuse them with a fluorescent reporter, such as GFP or mCherry...through detecting the reporter's fluorescence. Another way to label proteins is through treating cells with...distinguish different target proteins in your flow analysis, you'll need to use fluorophores that attach...mCherry, through genetic modification. As your protein of interest will only occur in combination with that...cells. The combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog Post...About Other Protein Tags Catch Up on Your Fluorescent Protein Background Which Fluorescent Protein Should ...long live chemistry! The discovery of green fluorescent protein (GFP) sparked a renaissance in biological...low fluorescence when free in solution, but high fluorescence upon binding to its cognate protein. In ...improvement of fluorescent proteins makes them the go-to choice for many (if not most) fluorescence microscopy...fusions on their own. Subsequent advances in fluorescent proteins have replicated many of the properties once...photon budgets are a widespread issue with fluorescent proteins—even the thriftiest cell biologist can feel...substantially brighter and more photostable than fluorescent proteins, providing a straightforward way to improve... -
Fluorescent Proteins 101: Aptamer Fluorophores
TypeBlog Post...oligonucleotides in fluorescent visualization techniques for decades, most notably for fluorescent in situ hybridization...applications including aptamer based fluorescent biosensors. Aptabeacons Fluorescence resonance energy transfer ...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...oligonucleotides over proteins is their modularity, which is lost as soon as the aptamer is bound to a protein fluor....that bound the compound malachite green (MG) to create a non-protein based visualization technique. Ultimately... be used to induce fluorescence outside the context of a paired fluorophore protein. Years later, Samie... -
Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP
TypeBlog Post... Cre-lox Resource Guide Fluorescent Protein Resource Guide Fluorescent Protein Blog Posts ...Other proteins may also be utliized as scaffolds, including the set of red fluorescent proteins. The T-DOGGs... similarly to GFP, but commonly used red fluorescent proteins dsRed, mCherry, and TdTomato do not induce...and derivatives GFP and YFP, but not red fluorescent proteins, as seen with T-DDOGs. Like T-DDOGs, Cre-DOG...system is likely applicable to many types of proteins. For proteins with a modular structure, constructing ...., et al. “A nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene manipulation...Caussinus, E., Kanca, O., and Affolter, M. “Fluorescent fusion protein knockout mediated by anti-GFP nanobody... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog Post...applications of fluorescent proteins. These fusion proteins enable the observation of proteins in living cells...oligomerization of fluorescent proteins is a concern. The propensity of fluorescent proteins to oligomerize...Inserting a fluorescent protein within your protein of interest We have encountered several proteins for which...Addgene blog Read our fluorescent proteins blog posts Download the fluorescent proteins 101 eBook Read our... Browse the fluorescent protein collection Find empty backbones for fluorescent protein fusions Browse...University of Amsterdam). Tagging a protein of interest with a fluorescent protein to study its function is one...fusion proteins and highlight some aspects of their design. Protein size and shape matters The green fluorescent... -
Tips for Using FRET in Your Experiments
TypeBlog Post...fluorophores to proteins, pioneers of this technique were able to directly detect protein/protein interactions...Day RN, Davidson MW. Fluorescent proteins for FRET microscopy: monitoring protein interactions in living... CFP-YFP was the first to be used to study protein-protein interactions and several other pairs have been... can be used to create your choice of fluorescent fusion protein with your gene of interest: Plasmid... 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...Articles About FRET: Piston DW, Kremers GJ. Fluorescent protein FRET: the good, the bad and the ugly. Trends... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog Post...mCherry fluorescent protein followed by a stop codon and the coding sequence for a green fluorescent protein...of cell-free system, capable of producing fluorescent proteins and enzymes. Image from Moore et al., 2021...validated through monitoring the expression of fluorescent proteins and oxytetracycline enzymes. Next, this ...biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The reaction...libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline...messenger-RBP sgRNA Library targets human mRNA binding proteins with 10 guide RNAs per gene. The lab modified ... -
FlipGFP, a novel fluorescence protease reporter to study apoptosis
TypeBlog Post...Addgene blog Read our fluorescent proteins blog posts Download the fluorescent proteins 101 eBook Read our... Browse the fluorescent protein collection Find empty backbones for fluorescent protein fusions Browse...necessary for fluorescence. When these parts are separated from each other, the protein cannot fluoresce. When...caspase results in a change in fluorescence emission. FRET and fluorescence based reporters however have...normal antiparallel position and fluoresce. FlipCherry: Red fluorescence protease reporter To add to the... cleave proteins. This protease reporter based on sfCherry however did not initially fluoresce in HEK293... used in FlipGFP can be applied to other fluorescent proteins to create an array of multicolor reporters... -
MXS Chaining
TypeBlog Post...Plasmids 101: Green Fluorescent Protein Resources on Addgene.org Browse our Fluorescent Protein Collection...combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters is a useful... sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging...been used to produce complex fluorescent reporter constructs. These fluorescent reporters can be used to ... The modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals...constructs were created, each containing a unique fluorescent protein with minimal spectral overlap between them...Table 1). In each construct, 3 copies of the fluorescent protein were fused in-frame to a tethering partner... -
Deep Dive: Fixing and Permeabilizing for Immunofluorescence
TypeBlog Post...the folded protein (green region). Methanol dehydrates the protein, which can disrupt protein structure...treatment for immunofluorescence suppresses localization of some integral membrane proteins. Journal of ... (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction that covalently... polymerize and lose its ability to cross-link proteins. As such, you will get the best results if you...solvents can be quite harsh - lipids and soluble proteins can be lost during fixation with these chemicals...these chemicals can also alter the structure of proteins, which could disrupt your target epitope (Fig....antibody binding. Formaldehyde cross-links the protein (red lines), which maintains structural epitopes... -
Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards
TypeBlog Post... awards to early career scientists using fluorescent proteins in their research. These awards help early... in the development and dissemination of fluorescent protein tools. This year, we’ve made some changes...inhibits the neuron (via hM4D). She fused fluorescent protein tags to the receptors to monitor their localization...are co-expressed with a photoconvertible fluorescent protein mEOS4a. Cells containing a sensor that shows...-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...thiol-containing acceptors. These hydropersulfides modulate protein activity and have important roles during oxidative... -
Plasmids 101: Gene, enhancer, and promoter trapping
TypeBlog Post...way. For example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed...construct can be something phenotypic (such as a fluorescent protein or something species-specific) or a selectable... -
Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative
TypeBlog Post... 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...DNA. TALE proteins can bind specific DNA sequences which brings the attached fluorescent protein to the ...10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining dye Article contributed...contributed by Leah Schwiesow Calcineurin is a protein phosphatase that responds to rising intracellular... in the cell, calcineurin dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent...this vector is suitable for flow cytometry and fluorescence microscopy, they also show that this method ... -
Hot Plasmids and Viral Preps - July 2021
TypeBlog Post...SL-sensitive fluorophore (circularly permuted green fluorescent protein or cpGFP) to the petunia SL enzymatic ...relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity, ...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... Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating a SL-insensitive fluorophore...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...