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We narrowed to 46 results for: Free Use GFP

Showing: 21 - 40 of 46 results

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

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ...widely used to study gene expression in cells. But these reporters have limited use in vivo because they...genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient transfection ...higher) of BFP and GFP producing cells. After FACS, return the cells to doxycycline-free media and expand...bacteria The first-generation bacterial ARGs (bARGs), used in bacteria, expressed poorly at body temperature...tissue unless destructive ultrasound imaging was used. We set out to develop next-generation bARGs that...expand the cells. Use a subset of the cells to verify the mARG positivity, which can be observed under phase... not require a booster cassette, either, allowing us to reduce the genetic footprint yet again. This ...

  2. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ...the spectra of enhanced GFP (eGFP) and FITC, the latter being a commonly used fluorophore for antibody-conjugated...stains that react with free amines on and inside the cells and are thus typically used for distinguishing ...commonly used fluorophores. Each fluorophore has a defined emission maximum. Note, you can also use the tool...mentioned previously, you can also use fluorescent reporters, such as GFP, and fluorophore-conjugated antibodies...you cannot use them together in the same panel. Source: https://www.bdbiosciences.com/en-us/resources/...software (without using the actual FITC fluorophore in our panel) will allow us to detect the eGFP signal. We...the fluorophores used in our panel (Table 2). Source: https://www.bdbiosciences.com/en-us/resources/bd-spectrum-viewer...

  3. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...such as GFP (Paix et al., 2014). A further development of this approach reported in 2017 uses the direct...organisms which I use to study gene regulation during developmental processes, CRISPR/Cas9 allows us to delete...drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes...first approach – co-CRISPR from Craig Mello’s lab – used inactivation of the unc-22 gene as their selection...marker (Kim et al., 2014). Meanwhile Andy Fire’s lab used oligo-mediated knock-in of a dominant mutation, ...even more powerful following a report that one can use linear repair templates (ie. PCR-derived dsDNA) with...co-conversion approaches have the advantage of being used in any genetic background, but require variable ...

  4. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ... They can also be used in crosstalk-free three-color imaging with green and red GFP-like FPs (Shcherbakova.... Modern state-of-the-art NIR FPs can be used similar to GFP-like probes by supplying a single gene to... Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum is perfect for use in spectral...35 kDa for miRFPs, and 27 kDa for EGFP) and a possibility to be used as internal tags due to the proximity...near-infrared) (Qian et al., 2019).  NIR FPs are useful in vivo probes, due to low autofluorescence, low...cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric... endogenous BV.  When imaging NIR FPs, consider using light sources that produce enough light intensities...

  5. Sensing Neuronal Dopamine

    Type
    Blog Post
    Published
    July 30, 2019, 12:59 p.m.
    ...sensor was engineered from a GFP variant and a DA receptor such that GFP fluoresces when the receptor ...release.  Using dLight1 sensors in experiments The Tian lab used dLight1 both in brain slices and freely behaving...on a similar strategy: a GFP variant bound to a DA receptor, such that the GFP fluoresces upon DA-receptor... for mouse or human receptors, respectively). Using GRABDA sensors in experiments The Li lab used GRABDA...quantitation is desired (Figure 1). The probe perfuses liquid, which causes the passive diffusion of DA across the...released DA in mouse brain slices Visualization of DA release dynamics during locomotion in freely moving mice...GRABDA in drosophila, zebrafish, and mouse brain slices and freely moving mice to gather the following types...

  6. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    Published
    Jan. 16, 2024, 6:13 p.m.
    ...Marker Mouse Strain C-CHIME Knocks out a pair of genes pXPR_219 Violet-excited GFP (vex) ...assessing genes in defined contexts would be quite useful for interrogating these scenarios. X-CHIME We recently...knockout of a single gene pXPR_068 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 L-CHIME ...types (lineage-based) pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks...of genes sequentially pXPR_220 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME...embryonic lethal, hindering studies post embryogenesis. Using C-CHIME and S-CHIME, we discovered that double knockout..., a suite of four systems which utilizes custom mouse strains and lentiviral expression vectors (Table...

  7. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...available biosensors use a single GFP or CFP/YFP-derived FRET pair. If you need to free up crowded optical...this blog we will explore some of the most commonly used as well as newly developed calcium sensors. Selecting...calmodulin or troponin C). Their fluorescent readouts are used to monitor calcium concentrations and transients...there waiting for you. When selecting a sensor to use for your studies, you can start by asking ‘What is...changes. The GCaMP6 family of sensors are commonly used calcium reporters with different kinetic parameters...experiment could be subtle or dramatic. You want to use a calcium sensor that is most sensitive to your expected...available. Detection methods Many calcium reporters use a single fluorescent protein (FP), requiring only...

  8. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...Ready to get started?   Figure 1: Break free from the ball and chain!   Epigenetics nation...Addgene has several great options: pLV hUbC-dCas9-T2A-GFP for removable dCas9 expression and pLM-CMV-R-Cre....sorting, and stable gRNA expression can be achieved using lentiviral backbones expressing blasticidin or puromycin...et al. (2016) Targeted DNA demethylation in vivo using dCas9–peptide repeat and scFv–TET1 catalytic domain...clearly testing whether a certain methylation state causes a condition. To understand mechanisms and, eventually...notoriety – as a tool of choice in the field: dCas9 fused to the enzymatic domain of TET1 or TET2 (Morita ... DNA-binding protein with no enzymatic activity cause demethylation?  Dividing cells depend on DNA methyltransferase...

  9. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...ambient light. In contrast, the widely used green fluorescent protein (GFP) gives a bright green fluorescence...directly use in bacteria, where most genetic manipulations start. Anthony Forster’s lab used codon optimization...anemone's red color can be used for mimicry. Image: Bernard Spragg. Advantages of using chromoproteins for ...for bacterial synthetic biology Because the common chromoproteins used in the lab come from eukaryotes...fluorescent proteins and luminescence have been widely used in biological research for a long time, chromoproteins...anemone, scientists think that chromoproteins could be used for mimicry and camouflage. An anemone’s red colored... it gives scientists the ability for instrument-free detection. Unlike fluorescence or luminescence, which...

  10. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...most commonly used CRISPR enzymes - Cas13a contains two HEPN domains, whereas Cas9 uses HNH and RuvC domains...starters, Cas13a can be used to bind and cleave target RNAs, although its usefulness in bacterial cells will...LwaCas13a), using a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can ...cleave any RNAs after binding a user-defined RNA sequence could be used to detect single molecules of an...exceptional genome engineering tool. It is easy to use, functional in most species, and has many applications...targeting with a single crRNA The Feng Zhang lab used Cas13a from Leptotrichia shahii (LshCas13a) to begin...2016). In the endogenous CRISPR/Cas9 system, Cas9 uses both a tracrRNA and crRNA to facilitate binding ...

  11. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...cell lines deposited with them and used by different researchers are free of contamination and match their... profiling can be used to ID the cell line. The same test is so good that it is used as a forensic technique...working vials have been used up. This ensures you can retain the maximum stocks and use of the original Cell...for Mammalian Cells Learn about the Dangers of Using GFP for Protein Localization Resources on Addgene.org...the present day Cell culture is a widespread tool used in the fields of oncology, virology, immunology,...specific to their origins. This technique is still used today to test for HeLa contamination. HeLa contamination... world has a unique STR profile) and can also be used as a paternity test. The test is accurate, precise...

  12. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...proteins in these compartments using CRISPR systems should strongly consider using moxFPs over standard FPs ...Generally, FPs evolved in or were engineered for use in cytoplasmic environments. However, approximately...green, cyan, yellow, and blue variants that can be used in various combinations. For example, with standard...herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion proteins in cell biology."Current...: PMC2784028. 3. Munro, Sean, and H. R. Pelham. "Use of peptide tagging to detect proteins expressed from...Lindsey M. Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted...Suzuki, Takahisa, et al. "Development of cysteine-free fluorescent proteins for the oxidative environment...

  13. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...method through monitoring GFP degradation, this protein compartment was used to optimize D-glucaric acid...population.  Because indel generation by Cas9 can occur at a steady rate, scientists found a way to use the accumulation... hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance... hope they'll make it easier for you to find and use the plasmids you need. Here's what you'll find in...bioproduction pathways The CRISPR corner New ready-to-use viral preps Mapping antibody escape in SARS-CoV...displayed on the yeast cell’s surface. The Bloom lab used the library to identify RBD mutations which do not...diseases, so these validated antibody tools could be used to study the role of aaRSs in these processes. To...

  14. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    Published
    March 10, 2015, 2:25 p.m.
    ...monoclonal antibodies to be used as a powerful reagent in the laboratory.   The DSHB focuses on monoclonal antibodies...Studies Hybridoma Bank. Antibodies are the most widely used class of protein-binding reagents. In the lab, their...antibodies against popular fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have...of generating antibodies, for example through the use of less toxic adjuvants; 2) generating complex antibody...antigens on foreign invaders, such as bacteria or viruses, which results in neutralization or targeted destruction...monoclonal antibody cell line, myeloma cells are fused with activated B cells from the spleen of mice injected...Technologies in Addgene's First 10 Years Data Freedom: The Expansion of Data Sharing in Research Publications...

  15. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    Published
    May 16, 2018, 1 p.m.
    ...That page has been very useful over the years, and when researchers come to us for advice on choosing ...soon as possible after submission. Using FPbase The most obvious use case for FPbase is finding information...designed with the goal of encouraging user contributions (using simple web forms) while also maintaining...contributions). Of course, no community-editable database is free from the risk of inaccurate data submissions, so...addition, the FPbase Fluorescence Spectra Viewer can be used to compare excitation and emission spectra when ...collections (such as a set of FPs that you or your lab use frequently) for the sake of comparison or ease of...can be kept private, or shared publicly (so feel free to moderate your own version of fpvis.org with the...

  16. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...tag often used for protein purification is a reliable FRET acceptor labeling site when used in conjunction... with reagents used to fluorescently label antibodies, and purification is a snap using thin layer chromatography...covalent, so free dye can be washed away. In addition, labeling contrast can be improved using disulfide-...finesse using tRNA suppression technology (Fig. 1) (Dumas, Lercher, Spicer, & Davis, 2015). By using a novel...Figure 2). These labeling methods are freely interchangeable and new uses for FRET are continually being developed..., depending on the probes). In fact, FRET can be used to derive a precise physical distance between the...organelles or other cellular impediments. FRET has been used in many applications, including drug discovery by...

  17. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    Published
    July 29, 2014, 6:16 p.m.
    .... In the world of fluorescent proteins and their use for imaging cell biology, Michael Davidson’s lab ...from Joachim Goedhart: Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection.  ... your protein of interest localizes in the cell, use these markers as your guide. As Michael Davidson ... we could, and Addgene is a wonderful vehicle for us to do that.” At Addgene, we are equally excited to...sun, from the standard oldies but goodies (e.g., EGFP and YFP) to the new and improved fruit colors (e.g...! Baird has a helpful tip for anyone looking to peruse the collection without getting overwhelmed by its...what we had made,” Baird said. “It won’t sit in a freezer at FSU. Mike is really the one who had been very...

  18. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ... hope they'll make it easier for you to find and use the plasmids you need. Here’s what you’ll find in...detecting strigolactones The CRISPR Corner New ready-to-use viral preps New toolkit for CRISPR-based genome...) Lentiviral vector design. B) Strategy for leak-free transcriptional control. C) Iterative transduction...enrichment of successfully targeted cells that can be used to generate organoid cultures with a modification...interest. While the authors optimized the workflow for use in a fetal-lung derived organoid system, the approach... strigolactones By Fabienne Furt Current methods used to detect strigolactones (SLs) are costly, non quantitative...germination of parasitic plants, this approach was also used to build and validate a ratiometric fluorescent ...

  19. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...Tips for using the agarose cloning method Test your agarose temperature. Cloning cells using an agarose... clones picked using the agarose method do not differ grossly from those isolated using cloning rings ...protein is fused with a fluorescent protein), fluorescence microscopy can theoretically be used to select...picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning...: After obtaining antibiotic-resistant colonies, use a Sharpie to circle 12-48 individual colonies by ...sterile tube (for 60 mm dishes only 1.5 ml of each are used). This solution is capped, swirled and tested for...well-spaced clones. For rapidly growing cells we sometimes use TPP peelaway T-75 flasks to provide a greater surface...

  20. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    Published
    June 15, 2017, 2:30 p.m.
    ...guarantee that using this fluorescent protein to tag your protein of interest will be problem-free. In figure...applications in which the fluorescent protein is used as an inert protein module. This is explained in...here. There are a variety of methods that can be used to measure your fluorescent protein’s propensity...ultracentrifugation yields an affinity and has been used to demonstrate that yellow fluorescent protein homodimerization...methods (based on fluorescence polarization) can be used to detect homodimerization. However, all the aforementioned...have developed a variety of plasmids that can be used in the OSER assay including CytERM-mKOkappa, CytERM-mTurquoise2...propensity to homodimerize. Several studies have used the OSER assay to demonstrate monomeric behaviour...
Showing: 21 - 40 of 46 results