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We narrowed to 369 results for: epor

Showing: 41 - 60 of 369 results

  1. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged... The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...couples NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected/expressing...NanoLuc luciferase. Both GpNLuc and OgNLuc BRET reporters produced bright signals without exogenous excitation...near 600 nm – a clear demonstration of why a BRET reporter can improve deep tissue imaging. The constructs...NLuc) and set about to create an in vivo imaging reporter by linking two CyOFP1 molecules to NLuc (CyOFP1...

  2. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    ... registered reports, the gold standard for testing hypotheses Finally, Registered Reports combine all ...Reproducibility Project: Cancer Biology, where reported methods may not include how many tests were conducted...attention on the results instead of the methods. Unreported flexibility and questionable research practices...analysis code, and other materials. The use of reporting guidelines can also help remind us to include ...important times to measure that growth rate, you report the results of all four of those tests and not ...fifth time has unpredictably high rates, should be reported as a new discovery that deserves confirmation ...best practices of Open Science. With a Registered Report, peer review occurs before results are known. The...

  3. The Addgene Antibody Data Hub

    Type
    Blog Post
    ...Antibody Data Hub. Data reports  Each antibody submission is used to create a data report, which contains experimental...that information in the report as well.    Figure 3: An antibody data report, showing the results of ...methods section for the data report shown in Figure 3.   At the end of the report, you’ll find a results section... the data report shown in Figure 3. The antibody is rated "Pass". Curating the data reports Just like... page of the Addgene Data Hub.   Antibody data reports can be searched and sorted by gene/target, name...information is then formatted and published as a report with a DOI for citation. If you’re considering ...

  4. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered using AAV injection...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...shown in Figure 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation...the two pairs of target sites. By flanking the reporter with one loxP site and one lox511 site in the ...would ensure that the reporter remains silent until a cell or tissue-specific Cre is provided. Since the FLEx...

  5. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as... screens to introduce reporters throughout an organism’s genome. These reporters have historically been...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...promoter – all rely on the random integration of a reporter construct in close proximity to an endogenous ...example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys... for with an uncharacterized gene. The actual reporter construct can be something phenotypic (such as...promoter. The system is composed of a promoter-less reporter which needs to integrate within an exon of a gene...

  6. Brain Initiative Collection

    Type
    Collection
    ...plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent expression Edward Boyden 108912...128485-AAV9 pAAV-CAG-iSeroSnFR-Nlgn Fluorescent reporter for serotonin (dendrite localization tag) Lin ...

  7. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ... transformation. Choose good reporter genes There are many great reporter genes available and which one...the T-DNA region with our gene of interest or a reporter gene so that the transformation and gene expression...different treatments on transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase...histochemical GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate...transformation success. Luciferases are great quantitative reporters and use a substrate to produce bioluminescence...half-lives then GUS and are therefore good temporal reporters to measure promoter activity, for example. Useful...Agrobacterium. This is especially problematic with reporter genes as you might observe expression from Agrobacterium...

  8. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ... versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27...The K9 reporter uses the HP1 chromodomain to recognise lysine 9 methylation, while the K27 reporter uses... methylation in living cells. Essentially this reporter construct consists of one substrate (the histone-derived...-K27 histone methylation reporter) positions on histone H3, which are known to be involved in repression...methylation of these sites was controlled, the reporter consistently showed significantly different FRET...states. Thus, this system is a reliable tool in reporting methylation status of these sites. This new technology...Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in living cells." Journal...

  9. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...and N177F mutations). Its excitation maximum is reported as 603 nm, while the emission maximum is 675 nm... ACE system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing and Cas9...editing tools, the Reuben Harris lab has deposited a reporter plasmid that allows for rapid fluorescence-based... out a frame-shift mutation within the mCherry reporter. Quantification of editing efficiencies simply...containing rat APOBEC1. Finally, they show that the ACE reporter can also quantify Cas9 cleavage, thus exhibiting...editing activities. Click here to find the ACE reporter plasmid! Martin et al., Nucleic Acids Research...

  10. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...random variation, or noise, in reporter expression. To do so, one places reporter expression under the control...expression of a reporter protein, it is possible to change the amount of noise in reporter gene expression...deliver the target antibody. TRIM-Away reagents and reporters are available from Addgene - we hope they will...lab has deposited plasmids encoding fluorescent reporters and other proteins controlled by GAL4 and ZIF268...progesterone controlled transcription factor as well as reporters specific to each transcription factor. They then... of estradiol and progesterone and showed that reporter expression is dependent only upon its cognate ...

  11. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...fluorescent reporter, directly couple silencing of your gene target to silencing of the reporter. This is...fusing your gene target sequence to a fluorescent reporter sequence (discussed below). Examples of these ...contains a target sequence against the fluorescent reporter in an inverted repeat orientation. Therefore, ...sequence against your gene(s) of interest and the reporter. This approach greatly enhances experimental throughput... observable phenotypes quantified. Fluorescent reporter-based RNAi constructs have been designed and widely...processes to isolate RNAi plants. With a visual reporter of RNAi, phenotypes, such as morphological parameters...identified as undergoing RNAi through the fluorescent reporter and manually extracted. This process is tedious...

  12. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...unique skills and strengths (you will likely have reports with different levels of experience), find out ...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...itself is counter-productive. I always assume that reports want to succeed at work and providing feedback ...feedback is needed for that. In many cases reports are not aware of issues until you bring them up. I wouldn’...wouldn’t go as far as saying that your reports will be grateful for the feedback, but at least you can start...someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk to your manager...

  13. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ... using IRES elements. Further, scientists have reported lower expression of the downstream cistron due...eukaryotic cells, and, although some scientists report close to 100% cleavage with some of these peptides...that already has the multicistronic element and reporter cloned in. In these plasmids you would simply ... 2A element (depending on the placement of the reporter gene). Addgene's collection offers a variety of...multicistronic element(s), or you could replace one of the reporter genes above with your gene or genes of interest...equivalent levels upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's...

  14. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...combination with that reporter, you can infer its presence through detecting the reporter's fluorescence. Another...labeling proteins is to fuse them with a fluorescent reporter, such as GFP or mCherry, through genetic modification...combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow cytometry...mentioned previously, you can also use fluorescent reporters, such as GFP, and fluorophore-conjugated antibodies...compare the excitation/emission spectra of your reporter with the ones from your fluorophore labels and...encodes for eGFP, which serves as a transduction reporter. Note that eGFP is not fused to CD45, but is separately...for having a separately expressed transduction reporter is that not every transduced cell will necessarily...

  15. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ... people usually report? Addgene and other viral vector manufacturing facilities report the physical titer...cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-infectious ... However, for recombinant AAV2, the same study reported a physical-to-infectious particle ratio of 50:...(“full”) (Zeltner et al., 2010), while another reported that only 20% of recombinant AAV2 particles were...vector preparation protocol, and we are pleased to report that over 95% of viral capsids in representative...AAV aliquots. Our titer values (circled in red) report physical titer, which is measured by qPCR. ...nonsense to publication quality data." Scientific Reports 7 (2017). PubMed PMID: 28546538. PubMed Central...

  16. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...mutagenesis libraries or to insert heterologous genes, reporters, or other markers into a target genome. They did...Rep 2014 Jun 27;4:5469. MitoTimer plasmids to report mitochondrial turnover Mitochondrial health...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to... to show its usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer... Yan’s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial ...channelrhodopsins. CnChR1, nicknamed Chrimson, is the first reported yellow-peaked channelrhodopsin with a spectral... original APEX (enhanced ascorbate peroxidase) reporter from Alice Ting’s lab has enabled progress on ...

  17. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since...fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was later found to be a...E., Sivanesan, D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein...Filonov, G.S. & Verkhusha, V.V. A near-infrared BiFC reporter for in vivo imaging of protein-protein interactions...evolution approach applied to fluorescent voltage reporters. Nat Chem Biol 14, 352-360 (2018). Yu, D. et al...

  18. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...luciferases, which have become common genetic reporters and alternatives to fluorescent proteins. ...Created with BioRender.com. Bioluminescent reporters require a chemical substrate (and sometimes cofactors...signal sensitivity. Another advantage: luciferase reporters aren’t susceptible to photobleaching — they’ll...firefly luciferase was the first bioluminescent reporter to be cloned and remains popular, other luciferases... for plasmids expressing luciferase Luciferase reporter assays for monitoring gene regulation Since firefly...approach has even been used in biosensors where BRET reports on the change in conformation of a single protein...expression vectors, and literally thousands of reporter constructs with a gene of interest already inserted...

  19. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed in ... and a reporter construct into the mouse retina. GFP expression robustly activated the reporter gene TdTomato...and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental manipulations...GFP. They then tested T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells ...tested in retinal electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, ...

  20. Newly Updated AAV Data Hub!

    Type
    Blog Post
    ...searching the Data Hub's curated collection of reports (which you can filter by serotype, species, or ... significant time and money. For instance, this report shows that anterograde, monosynaptic connections...our AAV Data Hub Challenge recording to see two reports presented by the submitting scientists, learn more...replicates of your experiment* Functional data, if the report describes a functional vector such as a channelrhodopsin...injection site as well as the target region, if the report describes a retrograde serotype An abstract that...experiment clearly and concisely* *required for a DOI. Reports that meet our minimum standards (* above) will ...
Showing: 41 - 60 of 369 results