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We narrowed to 361 results for: reporter

Showing: 21 - 40 of 361 results

  1. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...proteins to allow translation to occur. For a TRICK reporter mRNA this displacement leads to a change in the...GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP...translation, both GFP and RFP are bound to the reporter, causing it to appear yellow. The act of translation...of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization signal...copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors cycloheximide...is necessary for removal of GFP from the TRICK reporter. Delayed translation is known to be important ...

  2. Zinc Finger Consortium: Zinc Finger Arrays

    Type
    Collection
    ...Endogenous Zebrafish Gene Targets Recent published reports have shown that engineered zinc finger nucleases...greater in a well-characterized bacterial two-hybrid reporter system but have NOT yet been tested for activity...

  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. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...detectable by two different tags: a reporter and an HA tag. The reporter comes in three varieties: HRP, Spaghetti...independent of the reporter. The only caveat is that HA staining is weaker than reporter staining. Picking...Picking the right reporter for your RANBody While all three reporters stain cultured cells and tissue sections...Monster, and Chicken IgY Domain RANbody Reporters. Reporter Detection Method Uses Pros Cons Antigens...genetically tagged and instead are conjugated to reporters (i.e. fluorescent dyes, HRP, etc. ) after purification... is a RANbody? A RANbody is a fusion between a reporter and a nanobody that’s encoded on a plasmid. A ...sections, some reporters are better suited for particular applications. Read on to learn more about each...

  5. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...Fluorecent Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves... is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a...several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays...available to choose from) is now a common feature in reporter plasmids.  Learn How Luciferase Can Be Used In...transcription within cells, where detection of the reporter protein or its enzymatic activity is measured....expression can be determined by detection of the reporter in a specific assay, which ideally would have .... Production of photons by this bioluminescent reporter occurs slower than fluorescent-based methods, ...

  6. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI...Gal4 activator, bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including...to have Y2H systems that avoid transcriptional reporters altogether; In the split-luciferase system, intein-mediated...

  7. 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...

  8. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ...conjugated with a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such...dish and incubate it with a reporter conjugated primary antibody. The reporter reacts with a provided substrate...simplicity we will discuss the more common enzyme-based reporter methods. For enzymatic signal detection, users... sandwich and competitive. In a direct ELISA a reporter enzyme conjugated antibody binds to antigen coated...antibody first binds to the antigen and then a reporter enzyme conjugated secondary antibody binds to ...detection antibody can be directly conjugated with a reporter enzyme or a conjugated secondary antibody can ...).  The sample (purple) is preincubated with a reporter enzyme labeled antibody and then added to the ...

  9. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg... termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK and... SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was first identified in...molecules. In SHERLOCK a quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable...amplification step in combination with the ssRNA reporter enables SHERLOCK to detect DNA or RNA with atomolar...

  10. 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...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...notoriously difficult to publish.  With Registered Reports, the authors are able to make that clear distinction...

  11. 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...

  12. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ...set of lentiviral transfer vectors you can find reporter plasmids to test whether or not you've efficiently...to generate adenovirus expressing a variety of reporters as well as the Cre and Flp recombinases. Genes...

  13. 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 ...

  14. 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...

  15. 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...

  16. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    ...Career Development; a Stakeholder Workshop Report.” The report enabled the development of priorities for...Development Hub Workshop Attendees and their 2021 report. Image from pd|hub.   To begin to tackle these...the findings were captured in this just released report “Enhancing Dissemination of Evidence-Based Models...programs.  Please join us for the pd|hub Workshop Report Public Release Event scheduled for March 4th from...2:30-4:00 pm ET.  You can learn more about the report's recommendations and join with us to discuss and...

  17. 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...

  18. 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...

  19. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ...pyralis) reporter plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system...widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been...protein function, thereby creating a single-molecule reporter of intracellular processes. Multimerization via...the cognate att sites are supplied as individual reporter constructs or as the fully assembled pMemoryArray...

  20. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed... it is an attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics... target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay, and are...Büttner’s lab designed a calcineurin transcriptional reporter consisting of a quickly-degraded destabilized ...panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate ...Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial...
Showing: 21 - 40 of 361 results