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We narrowed to 62 results for: Free Use GFP

Showing: 41 - 60 of 62 results

  1. Genetic Code Expansion

    Type
    Collection
    ...first with a control reporter gene – GFP for E. coli or mCherry-GFP for mammalian cells. You should also...to be allocated to the new amino acid. Because there are no free codons, this can be challenging. In E....Addgene's guide to plasmids and strains useful for expanding the genetic code in bacteria and mammalian...amber stop codon (UAG) and thus this codon is often used. The gene of interest can be expressed from a plasmid... observed. The introduced amino acid can also be used to intentionally change the activity of a protein...growth medium, ncAA concentration, and the cell lines used previously. Remember that the orthogonal pairs of...unallocated codon. Therefore many controls must be used to make sure that these conditions are true. Always...

  2. Adenovirus Guide

    Type
    Guide
    ...limited their use in therapeutic applications. However, this has also made them the most widely used viral vector...pAdTrack series contains an IRES-GFP construct that enables co-expression of GFP with the transgene of interest...viral vector production. During experiments, GFP can be used to sort cells infected with rAdV vectors, or...based on the use of pAdEasy-1. Created with BioRender.com. AdMax™ AdMax™ is another well used rAdV vector...disadvantage for the use of rAdV vectors in gene therapy but has, however, proven to be very useful for designing...that contain IRES-GFP. Pseudotyping The production of viral vectors (or viruses) using viral envelope proteins...that do not contain GFP. Recombinant adenoviral vector (rAdV) Modified form of AdV used to deliver genetic...

  3. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...FlincG3 (GFP-based cGMP sensor) for imaging in C. elegans neurons Using a Robust and Sensitive GFP-Based ...negative-control constructs used to characterize the sensor's behavior in the target system. Use care when applying...Calcium erGAP3 (GFP-Aequorin Protein) for imaging of Ca+ dynamics in endoplasmic reticulum GFP-Aequorin Protein...browse all the biosensors tagged in our catalog . Use the article links to find the plasmids associated...associated with the article. We also offer ready-to-use AAV preparations of select plasmids and AAV Packaged on...2020 GENIE Project Calcium jGCaMP7 High-performance GFP-based calcium indicators (Constitutive or Cre-dependent...Fluorescent Mg2+ sensor (MARIO) A Transient Rise in Free Mg(2+) Ions Released from ATP-Mg Hydrolysis Contributes...

  4. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  6. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...Production Using Packaging Cell Lines Gamma-retroviral vectors can be packaged using helper-free cell lines...Common Uses of Gamma-Retroviral Vectors Due to their integration, gamma-retroviral vectors are useful research...-activated cell sorting (FACS) is used to sort cells expressing GFP and later, sorted cells are expanded...clinical studies and commercial purposes. The use of helper-free production cell lines also saves time in ...viral vectors (or viruses) using viral envelope proteins from different species. Used to alter infectivity...variety of cell types than retroviruses. Both lentiviruses and gamma-retroviruses use the same packaging genes...Pseudotyping Common Uses Safety Resources Plasmid Elements Glossary Gamma-retroviruses are a subtype of ...

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

    Type
    Blog Post
    ...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...

  8. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    .... 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...

  10. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ... 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 ...

  11. Antibody Production

    Type
    Collection
    ...antigen is not endogenously-expressed (e.g., using an anti-GFP antibody), the target antigen is first transiently...not recommend repeated freeze thaw of antibodies and suggests preparing single-use aliquots before storing...Addgene have been produced in-house by Addgene scientists. They are generated using standard methods individually... sodium azide. This buffer is not compatible for use in live cells and will interfere with conjugation...our other resources to help you choose, store, or use your antibody. Antibody Blog Posts Recipient Instructions...for 7 days and then purified from the supernatant using Protein A or Protein G affinity columns. Formulation...chromatography, antibodies undergo a buffer exchange using centrifugal columns. The final formulation buffer...

  12. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  14. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...mammalian cells. Their marker-free coselection, analogous to methods previously used in C. elegans, targets ...can be used to measure the size of DNA fragments. Depending on which restriction enzyme is used to cut...mutants), gRNAs used for TZAP gene editing, and a Hi6xs-MBP tagged TZAPznf9-11constructs used for bacterial...power of mCyFP1, the Lin and Yasuda groups use both mCyFP1 and EGFP as FRET donors in experiments simultaneously...popular CRISPR base editing technique uses dCas9/Cas9 nickase fused to a cytidine deaminase for targeted...these articles make it easier for you to find and use the plasmids you need. You can find all the hot plasmids...-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked...

  15. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...developed and used AdV vectors to edit genes in several biological systems. It has been used for instance...Addgene and could be useful to labs wishing to create this kind of animal models. Start using adenoviral vectors...Benoit Giquel.   Adenoviral vectors (AdVs) have been used for many years for in vivo delivery and gene therapy...donor DNA is copied into the cut site through error-free homologous recombination (HR). Adapted from an image...viral particle. The earliest explorations of the use of AdV for in vivo CRISPR-Cas delivery utilized the...homology directed repair and you would probably need to use a two viral vectors system: one vector for Cas9/sgRNA...Dystrophy, Alpha1-antitrypsin deficiency (most widely used so far) or for cancer gene therapy such as nonalcoholic...

  16. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...validated your gRNAs for use in mammalian cells, it’s time to use them in mouse embryos. When researchers... Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use in mammalian...CRISPR Software Matchmaker can be used to determine the pros and cons of using many different types of gRNA...weeks old and use the ear tag method (creating different patterns of ear holes and using that material...and fastest way to use the system in mice? We'll have a post that goes into the mouse genome editing process...efficacy in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully this ... of EGFP to recombine by homology directed repair, and resulting in the expression of EGFP. Using this...

  17. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ...tend to photobleach rapidly, which hinders their use for long-term imaging or advanced techniques like...delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis. Nature Plants, ...Recovery (LARRYv2) barcoding libraries. The team used these libraries to analyze pre-existing heterogeneity...tools in your experiment. These libraries will be useful for studying the impacts of other cancer mutations...Addgene has tested these capsid plasmids in vitro and uses them in our own AAV production processes. Check ...Can't wait? Browse our In-Stock AAV inventory or use our Packaged on Request service for thousands of ...eligible AAV plasmids to order viral vectors ready to use in your lab. Find Addgene’s AAV6 and AAV11 packaging...

  18. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ... by using a spreadsheet. The plasmid information can be entered using our online form or by using a downloadable...gRNAs are used for directing CRISPR/Cas9 to a targeted genomic location, while shRNAs are used to knockdown...that can be used under BSL1 conditions were generated in your lab are necessary for use of a plasmid...deposited with us and distribute them to scientists worldwide. What's more, depositing is free! We've written...Either choice can be used for deposits of less than 10 plasmids but we recommend using the spreadsheet option...IDs.   Name – we encourage the use of descriptive plasmid names and using the exact names listed in your...needed. Use the "Relevant mutations/deletions" box to describe any mutations within your insert using standard...

  19. Modular Cloning Guide

    Type
    Guide
    ...circuits using Golden Gate-based cloning methods for use in mammalian cells. These can be used with plasmids...nickase, dCas9, or dCas9-FokI fusion for use in human or mouse systems. Fragmid Genome Engineering, CRISPR...resulting in a 4-bp fusion site in between. MoClo can be used for a variety of applications—like building synthetic...cloning vectors (Level 0, 1, and 2) which can be used in successive assembly steps. Before beginning, ...standardized and characterized parts that can be used for bottom-up hierarchical assembly of single and...Yeast Expression Paul Young 34 plasmids that can be used in conjunction with the MoClo Yeast Toolkit (YTK...integration in fission yeast S. pombe . These can be used with plasmids from the MoClo-YTK . MoClo Pichia ...

  20. CRISPR Guide

    Type
    Collection
    ...these PAM-flexible Cas9s are useful for precision edits, they can be used for virtually any CRISPR application...concentration too low to be used in experiments and must be amplified before use (Figure 8C), following the...libraries have been developed for inhibition using dCas9. How Do You Use a CRISPR Library? CRISPR libraries from...co-opted for use in eukaryotic cells . The Cascade complex is recruited to target DNA using a gRNA and ...bacterial genomic region used in pathogen defense CRISPRa CRISPR Activation; using a dCas9 or dCas9-activator...multiple genes at once; using dual nickases to generate a knockout or gene edit; or using Cas9 to generate large...will look for relevant hits using NGS technology. For more detail on using CRISPR libraries for screens...
Showing: 41 - 60 of 62 results