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We narrowed to 155 results for: GFP expression plasmids

Showing: 41 - 60 of 155 results

  1. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...vectors are tested in vitro and in vivo for gene expression and/or function. These data are sometimes reported...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ...the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from ...

  2. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq PUF Assembly... of GFP and RFP fusions as their test genes, the researchers at the BIOFAB showed that expression of many...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...your polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the...Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved for both...benchmarking of cell-free expression systems, and tested for product integrity, expression level, and aggregation...

  3. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    Published
    Nov. 19, 2019, 2:08 p.m.
    ...activates Ras. They expressed this OptoNB-SOS fusion protein in cells that expressed membrane-localized... Resources on Addgene.org Find nanobody expression plasmids here Learn more about optogenetics in Addgene's...Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins...co-localization with a membrane-localized mCherry or GFP protein. With this system, if the OptoNB can bind...more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop 1 always generated....  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced dark-induced ...until the message reaches the nucleus where gene expression changes lead to cell proliferation and differentiation...

  4. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ...promoter is expressed. Cre-mediated deletion of one or several of these genes permits the expression of genes... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ...fluorescence of the downstream OFP, GFP, or RFP. Cells that express the construct but do not undergo recombination...activate the expression of a different fluorescent protein. Therefore, each cell could express a unique combination...this mouse generates cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic...copy of the transgene expressing either YFP or CFP, and cells that do not express Cre fluorescing with ...the low expression of XFP of Brainbow by incorporating a Tet-Off system to boost XFP expression. The increased...

  5. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ...Protein Activity with GFP Resources on Asddgene.org Find Fluorescent Protein Plasmids Find Fluorescent Protein...signal to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...the amount of background fluorescence from unbound GFP-dCas9. With these improvements, labeling efficiency...sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin binding ...hairpins generates white light. All 6 gRNAs can be expressed in a single vector, pCRISPRainbow-DONOR1, with...CRISPRainbow gRNA will permit labeling, but a newly expressed standard length gRNA would “switch” the system...24360272. PubMed Central PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui...

  6. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...Fluorescent Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of EGFP can lead to significant...contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies ...protein or Enhanced Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general... a tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is in a...in the world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently...best solution is to use truly monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP...

  7. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ...In the presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR.   Sometimes...elements to turn off expression of one gene, while simultaneously turning on the expression of another! What...FLEx switch that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to ...presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead of BFP. Beyond...allowing for expression of GeneR. This allows a researcher to test whether or not expressing GeneR alone...potential to express the transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic...ensures Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...

  8. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...

  9. Antibodies 101: Epitope Tags

    Type
    Blog Post
    Published
    May 11, 2023, 1:15 p.m.
    ...antibodies (mouse, rabbit, and chicken) to this tag! GFP  GFP is more than a peptide – it’s a 238 amino acid...localization. If you need monoclonal GFP antibodies or guidance on designing a GFP tag Addgene has you covered!...endogenous WT levels of the same protein when both are expressed in the same system. Tagging can also aid in protein...acid protein that can also be used as a tag! GFP was originally derived from jellyfish (Aequorea victoria...epitope tag as well as a fluorescent marker. The GFP tag is ideal for live cell imaging, FRET, flow cytometry...antibodies also exist for imaging. The obvious con of GFP is its size – which can possibly interfere with protein...Addgene blog Antibodies 101: Epitope availability Plasmids 101: Protein Tags Antibodies: Of Myc and Men ...

  10. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness... There are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made...examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and...its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles a cylinder...2013; Hilaire et al., 2017). GFP can only be used for tagging proteins GFP fusion proteins are made by ... a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP fluorescence overlaps... al., 2018). GFP is not tolerated in my fusion It is standard practice to attach the GFP to the N- or ...

  11. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ... in a plasmid with loxP sites such that their expression is turned on or off in cells expressing Cre. ...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes... to track where GFP labelled B cells were in mounted slices of these organs. These GFP labelled cells ... depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after ...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...enabled by fluorescent proteins The optimization of GFP and its derivatives by the late Roger Tsien changed...stimulus. Using an epifluorescence microscope and GFP fusion proteins controlled by promoters specific ...

  12. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...transduced at levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi....need. If you'd ever like to write about a recent plasmid deposit please sign up here.        AAV delivery... Figure 1: The base editor is split into two plasmids and co-transfected into cells for reconstitution...voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...

  13. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...transcriptional activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element.... Today, it's time for another edition of Hot Plasmids!  Here's what you'll find in this post:  New...., 2023 under CC-BY 4.0 License. Find PE6 plasmids here!  Doman, J.L., Pandey, S., et al. (2023)....therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific...pulse-chase experiments, and more. Find LOV-Turbo plasmids here! Next, Qin and Cheah et al. described TransitID...LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments open up new possibilities...

  14. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be identified...conjugative mobilization of broad host-range IncQ plasmids." Plasmid 62.2 (2009): 57-70. PubMed PMID: 19465049...page Deposit microbiology plasmids with Addgene Find synthetic biology plasmids ...family-RP4 conjugation system. This system can deliver plasmids to both Gram-positive and Gram-negative cells.... Wang lab further developed a modular suite of plasmids (pGT) with various replicative origins that can...selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both sorted using FACS and isolated...genera across four phyla that have taken up the plasmid through conjugation in vivo studies. However, they...

  15. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in an inactive...electron microscopy analysis is time consuming. GFP-tagged Atg8p or LC3: ATG8 is part of a group of genes...with autophagosomal membranes, so tagging it with GFP allows for tracking the localization or accumulation...autophagosomes and autophagic bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p homolog...pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic pHluorin (SEP). They are ...yeast Rosella is targeted to the cytosol when expressed without a signal sequence and to the mitochondria...Fluorescent Protein Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy...

  16. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent...AAV for expression in forebrain GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...too high expression of these markers can lead to non-specific labeling. They generated plasmids tagged ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...

  17. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    Published
    July 27, 2017, 1:11 p.m.
    ...lactose released activates expression of GFP from the lac operon. GFP expression can then be used to determine...fantastic tools it’s now easy to monitor protein expression, localization, and protein-protein interactions...(including lactose), and a lac operon controlled GFP. When the enzyme breaks down the HMO of interest,...Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids from the Tsien Lab Browse the Michael Davidson...second sensor is a fusion protein consisting of sfGFP fused to a His-tagged ABC transporter known to bind...

  18. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    .... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding...in patched cells expressing wild type CoChR, these APs are absent in soCoChR-expressing cells. Finally,...further probe cellular function. The CRY2clust Plasmid can be found here. Park et al. Nat Commun. 2017...C-terminus of the protein. Unlike the diffuse neuronal expression observed with wild type CoChR, soCoChR localizes...

  19. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ...constitutively-expressed mARG plasmids driven by CMV promoters. Labs that are new to the mARG expression are advised... that, when expressed, enable cells to be visualized under ultrasound imaging. Expression of ARGs results...in bacteria, expressed poorly at body temperature (37oC) and were too burdensome to express in situ. As...that the plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe...Fig. 1c).     Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe... gene expression in deep tissue. However, not unlike the first-generation bARGs, poor expression and acoustic...Increasing expression and decreasing genetic footprint We took a cue from the native expression patterns...

  20. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ... hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...indels to measure time. These plasmids with different promoters driving expression of Cas9 were used in this...vectors expressing cytoplasmic targeted tdTomato and pre-synaptic targeted synaptophysin tagged eGFP. AAV...Hutchinson Cancer Research Center developed a pooled plasmid library encompassing all possible amino acid mutations...
Showing: 41 - 60 of 155 results