...bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the viral...32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin Ng High-throughput screening...cell types expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP variant...splitting and joining the protein regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd...use light to induce conformational changes in a protein.    Cells expressing an RFP reporter plasmid...screening methods, such as fluorescence-activated cell sorting (FACS) helps researchers sift through thousands...Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand break repair assay...

...the core of the protein. OptoNBs were also tagged with an infrared fluorescent protein tag which let the...choose your own protein purification adventure. You want to purify your favorite protein (YFP). You have...bind and release a protein of interest in response to light allows for tagless protein purification as well...purification of unmodified proteins. Affinity purification methods typically require a protein to be tagged, which...approaches to use with recombinant proteins. Purifying an untagged protein with a flip of a light switch would...to light-controlled protein binding, they probably don’t bind your favorite protein yet. More OptoNBs would...binding.  Light controlled protein purification with OptoNBs The first application for OptoNBs takes us in...

...recent contribution to the field is a series of fluorescence resonance energy transfer (FRET)-based calcium...colleagues began by isolating a calcium-binding protein, troponin C (TnC), from muscle cells. They identified...calcium binding and developed a version of the protein with as few calcium-binding sites as possible for...transcription factor cAMP-responsive element-binding protein (CREB) in living cells. “Now we are making this...Eindhoven has similar interest in FRET-based sensor proteins, particularly for the intracellular imaging of...with a solution: they devised self-associating fluorescent domains whose association is disrupted in the...Robust red FRET sensors using self-associating fluorescent domains. ACS Chem Biol. 2013 Oct 18;8(10):2133...

...Which Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein...leash for your protein. If you’re working with an uncharacterized protein, or a protein for which a good... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the...Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used as tags for detection...fusion protein tags. Biochemists and molecular biologists who need to overexpress and purify proteins can...tags and fusion proteins to aid in the expression and purification of recombinant proteins. Tags for stability...recombinant protein? It is not generally in a cell’s best interest to overexpress a protein. Energy and...

...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...acids (DNA and RNA) absorb maximally at 260 nm. Proteins on the other hand absorb best at 280 nm and organic...sequencing. Unlike absorbance-based methods, fluorescence-based methods require a standard curve, a set...this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary electrophoresis...These fragments are quantified over time using a fluorescent dye that intercalates into the DNA such that ... using capillary electrophoresis which shows fluorescence units over time. Smaller fragments migrate more...

...MitoTimer proteins are derived from the Timer protein - a mutant of the fluorescent protein dsRed that...three possible fluorescent proteins, chosen for minimal spectral overlap, minimal protein aggregation, ...groups to monitor protein and cell ageing, as the spectrum shift occurs during Timer protein life due to a... synthesized (green) protein and organelles with older (yellow to orange) protein. Image courtesy of Roberta...1852-61. A quicker method for protein inactivation Your protein of interest does x, but it also participates...your endogenous protein of interest and co-expressing its replacement recombinant protein with a FKBP domain...outer membrane protein with a FRB domain (MitoTRAP). Rapamycin binds your recombinant protein (via FKBP) ...

...intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged with 10-24 copies...contains an HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just...which uses an scFv to amplify the fluorescence intensity of a tagged protein. Resources on Addgene.org: ...often genetically fused to other proteins, which has led to their application for basic and translational ... research applications of scFvs: The SunTag system uses an scFv to amplify the fluorescence intensity... are often the building blocks for engineering proteins. So let’s get acquainted with scFvs. What is an...antibody.  The ability to combine scFvs with other protein domains, much like lego pieces, makes them better...

...C terminus of a protein may disrupt the biological function of some proteins. Protein folding issues, ...of detection of a mutant protein vs. endogenous WT levels of the same protein when both are expressed ...to a protein’s termini to increase detection. The tag can also be removed by treating the protein with...validate tagged protein functionality (Arribere, et al). In rare cases, tags can also affect protein localization...of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol...antibody that works for your application, specifically detects your protein, is species compatible, and...other proteins, quality antibodies do not exist, or they don’t exist for a specific application, or they...

...colleagues identified a highly bright and stable fluorescent protein, StayGold. Although it quickly saw wide interest... E., et al. (2023). A monomeric StayGold fluorescent protein. Nature Biotechnology, 1–4. https://doi.org... al. (2023). Bright and stable monomeric fluorescent proteins derived from StayGold. ResearchSquare (Preprint...recombinant protein, the FLAG tag has proven itself a good option for essentially all applications. Addgene...Lab addressed this problem by optimizing the Cas protein delivery with Peptide-Assisted Genome Editing (...dimerization while maintaining the bright and stable fluorescence of the original (Ivorra-Molla et al., 2023)....mature marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human...

...recruit specific RNA-binding proteins (RBPs) tagged with fluorescent proteins to visualize targeted genomic...compatible with fluorescence microscopy in living cells. Alternatively, gRNAs can be fused to protein-interacting...catalytically inactive Cas9 (dCas9) fused to a fluorescent marker like GFP, researchers have turned dCas9...live cell imaging. Compared to techniques like fluorescence in situ hybridization (FISH), CRISPR imaging... the chromatin dynamics in living cells. The fluorescent CRISPR system has been used for dynamic tracking...requires recruitment of many copies of labeled proteins to the given region. For example, chromosome-specific...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...

...chimeric protein consisting of Cas9 and a photo-dissociable dimeric green fluorescent protein (pdDronpa...CRY2) and calcium and integrin-binding protein 1 (CIB1) proteins. The goal of both groups was to create...composed of two fusion proteins: 1) the genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1...form a functional protein due to being fused with high-affinity interacting proteins Coh2 and DocS, respectively... saCas9 protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly... architecture (Hoffman et al., 2020). Fluorescent dCas9 proteins can be used to identify specific genomic...telomere-targeting gRNA and a dCas9 fusion protein with three red fluorescent tags in U2OS cells. Upon light activation...

...expression system. Fluorescent Protein Guide : Addgene's extensive fluorescent protein plasmid collection...systems) and therefore supplied as one item. Fluorescent Proteins FRET-based biosensors : E. coli expression...three-part fusion proteins - Planas and Santalucia Labs pCri System : Heterologous protein expression in ...EcoFlex MoClo : Modular cloning for applications like recombinant protein purification and cell-free circuit...Kits for Microbiology : Sets of plasmids for fluorescence, cloning and expression, and synthetic biology...expression - Pertz Lab Bacterial Fluorescence Labeling : Broad-Range Rainbow Vectors - Barbier Lab Cloning and... : 285 vectors for expression, markers, fusion proteins, etc - Lindquist Lab Synthetic Biology BIOFAB ...

...plasmids/viruses are needed to deliver all the protein and RNA components, and several sgRNAs must be ... start sites, all of which yield the same BDNF protein. To demonstrate the specificity that can be achieved...The pooled nature of this experiment requires fluorescence activated cell sorting (FACS) to isolate differentiated...inserted human CD8, a gene that encodes a surface protein for which there are good flow cytometric antibodies...recent advancements and offer commentary on their application to neuroscience research. What is CRISPR-based...accessible to neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly...study neuronal differentiation, it could also be applied to the differentiation of other CNS cell types,...

...collections. Depositor Collection Types CRISPR Fluorescent Proteins Protein Expression Synthetic Biology Viral Vectors...pooled libraries. Find tools for CRISPR, fluorescent proteins, optogenetics, biosensors, and more. Addgene...Cre-lox TALENs Zinc Fingers Luminescence Fluorescent Proteins Luciferase Optogenetics Chemogenetics Viral.... Click here to view all kits. Kit Types Fluorescent Proteins Cloning Genome Engineering Pathways & ORFs...RESOLUTE Plasmid Collection for Solute Carrier Proteins (SLCs) Structural Genomics Consortium Zinc Finger...resources group plasmids together based on type or application. Browse our guides to find the plasmids and science...

...Alias: CN4496) Expression of SYFP2 (yellow fluorescent protein) plus separate nuclear FLAG epitope in striatal...Alias: CN2610) Expression of SYFP2 (yellow fluorescent protein) in striatal direct pathway medium spiny ...Alias: CN2408) Expression of SYFP2 (yellow fluorescent protein) in striatal and neocortical Sst-Chodl neurons...Alias: CN2787) Expression of SYFP2 (yellow fluorescent protein) in striatal Pvalb+ neurons. Note strong ...Alias: CN3038) Expression of SYFP2 (yellow fluorescent protein) in striatal and lateral septum cholinergic...Alias: CN2700) Expression of mTFP1 (cyan fluorescent protein) in striatal indirect pathway medium spiny...Alias: CN2237) Expression of SYFP2 (cyan fluorescent protein) in striatal indirect pathway medium spiny...

...cells in live imaging application. Each visual barcode consists of a fluorescent protein coupled to a specific...GFP-nanobody rather than a specific protein of interest. This fusion protein can then be co-expressed in essentially...stabilizes the TurboID-nanobody protein and allows TurboID to start labeling proteins in close proximity to the...localization peptides with four different fluorescent proteins, and inserting each visual barcode into ...reporters that drive the expression of the fluorescent protein by a specific transcription response element...approach has been hampered by the need to tag each protein of interest and, for in vivo work, generate a dedicated...zebrafish line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to...

...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs...imaging in C. elegans By Alyssa Neuhaus Red fluorescent proteins like mScarlet are essential tools for visualizing...under CC-BY license.  These optimized fluorescent proteins will be a useful addition to the toolkit... mScarlet3 for applications needing intense brightness (such as low-expression proteins of interest) and... produced by our partners at the Institute for Protein Innovation (IPI), part of a new collection of epitope...short, linear epitope derived from the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987)...including popular new approaches like PROTAC targeted protein degraders. But CRBN has been difficult to study...

...decorated with a fluorescent protein remain functional. Attaching the fluorescent protein fusions allowed...produced by orchestrated movements of networks of proteins consuming molecular fuel sources. While the importance... can be generated by conformational changes in proteins? How can information propagate through a structured...rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony explanation...addressed using R bodies: simple, force-generating protein machines that can extend and retract over many ...

... decade, but the fluorescent proteins (FPs) necessary for many of these applications have lagged. Fixation...Campbell, B.C., et al. Chemically stable fluorescent proteins for advanced microscopy. Nat Methods 19,...immunoassays due to only having one binding site per protein. Having multiple antibodies allows you to use them...light and electron microscopy and observed high fluorescence resilience to these preparations. After crystalizing...    Fig. 4: hfYFP retains folding and fluorescence in high molarity GdnHCl, at high temperatures...antibodies may just not perform as well in one application as another, so it is always best to have options...for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page for more...

...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...binding, changes in fluorescence can be visualised. Another approach was to apply the same concept to ...streptavidin portion of the probe via biotin. The fluorescent portion of the probe that is cut will continue...and released when cleaved. Cleavage results in fluorescence and this approach can be incorporated into a...test varies in the sequence of steps and the Cas protein used. Here, all the components needed for the reaction...recombinase, and the ssDNA-FQ reporters that exhibit fluorescence upon cleavage. Upon amplification by the RPA...