Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 902 results for: nes

Showing: 321 - 340 of 902 results

  1. Antibodies 101: Conjugation

    Type
    Blog Post
    ...either through a chromogenic or chemiluminescent reaction. Chemiluminescence, which is light produced by ...the primary amine groups from lysines, sulfhydryl groups from cysteines, or carbohydrates from glycosylation... found throughout the antibody. 2) Endogenous cysteines, such as those that make up the disulfide bonds...antibody (Mueller, et al., 1988). If some of those lysines are in the antigen binding region or even just ...

  2. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...into ‘split-vector systems’ by separating viral genes and transgene expression across several plasmids...that allow functional screening of thousands of genes in genome-wide experiments. Gamma-retroviral vs...direct the cellular RNAi machinery to silence target genes. This empty MuLV vector from the Weinberg lab enables...find more lentiviral expression vectors (empty backbones) here. Cloning and expressing new shRNA sequences...lentiviral backbone from the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette. It can be used to produce...studies. The ability to simultaneously alter multiple genes via infection with a single ecotropic or amphotropic...

  3. Hot Plasmids - October 2020

    Type
    Blog Post
    ...So how can you use CRISPR to introduce large FP genes into C. elegans? By using a split FP approach. Split-wrmScarlet... fluoresce. The team engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other... validated this tool by targeting six different genes whose protein products have distinct localizations... library that targets human glycosyltransferase genes and other proteins related to cellular glycosylation...deletes human alternative cassette exons and disrupts genes with alternative exons. Get the library from Addgene... format! The library inhibits over 18,000 human genes. Find the lentiviral preps at Addgene hSyn1-SIO-stGtACR1...

  4. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...stripping to remove, so start with the weaker binding ones first to preserve your sample. Pro tip! Because ...reprobing works best with western blots that use chemiluminescent or fluorescent detection, like horseradish...protein retention. Following this step, most PVDF membranes recommend re-wetting with methanol before rinsing...the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed with caution. Unlike PVDF...your own stripping solution using a recipe like the ones shared by Bio-Techne, ThermoFisher, or Millipore...remains, whether that involves adding fresh chemiluminescent substrate or imaging your fluorescent probe...

  5. CRISPR in the Clinic

    Type
    Blog Post
    ... to, ones targeting muscular dystrophy, cardiovascular disease, inflammatory diseases, blindness, and ... cancer treatment by traditional gene therapy pipelines (viral), and researchers are currently working...many more in the earlier stages of the pipeline, ones that have experienced hiccups (either clinically...challenges with therapies involve patient safety, effectiveness, and delivery. Patient safety is important for...concerning when using a systemic approach. Regarding effectiveness, if the editing frequency isn’t high enough ...

  6. Hot Plasmids February 2024

    Type
    Blog Post
    ...expression yielded editing of 80–90% in human cell lines and primary cells. Although cell entry of a Cas9...minimal toxicity, including in clinically-relevant lines such as CAR-T cells. Plus, this method is fast! ...break up the dimerization without losing that brightness and stability. Here’s a quick overview of these...variants:   Figure 2: Inner mitochondrial membranes imaged with structured illumination microscopy.... This variant has similar photostability and brightness compared to the original, though its maturation...versions. Thanks to Christophe Leterrier, whose FocalPlanes piece on mStayGold inspired and informed this...

  7. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ... constructs for tagging the kinesins KIF5A and KIF5B as well as backbones for generating other SNAPf-,...the conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling... will be useful not only for future studies of kinesins but for many other proteins of interest too! Find....A.S., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation...mouse monoclonal antibodies targeting the human genes and are recommended for use in immunohistochemistry...

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

    Type
    Blog Post
    ...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding...Homology Directed Repair (HDR) to knock in corrective genes. For this you would need to provide a DNA template...successfully developed and used AdV vectors to edit genes in several biological systems. It has been used ...plasmids at Addgene: pAdSh.PGK.Cas9 (expresses S. pyogenes Cas9 from the PGK promoter) and U6 promoter-driven...CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4:...

  9. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ... (24-30°C). Brightness: Brightness is a measure of how bright is the emission. Brightness is calculated...contributed by Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how fluorescent proteins glow...the protein, divided by 1000. In many cases the brightness is compared to that of EGFP which is set as 1...proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be taken into account...

  10. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...namely that they can cause cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular... are, in most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet ... is (thankfully) gutted of many essential viral genes. How it is, exactly, that heaps of viral vector ...the only component that is not dependent on viral genes. It is derived from the host/producer cell membrane...full “life cycle” of expression of selfish, lazy genes of a non-living viral particle, and how we as dedicated... N. (1971). Noninfectious ARK mouse embryo cell lines in which each cell has the capacity to be activated...

  11. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...changing guidelines and regulations impacting grant management, we track grant opportunity deadlines, and ...organization to evaluate the organization’s grant readiness, identify grant opportunities, secure grant awards...schools, healthcare facilities and with private businesses. How do you get the expertise needed to enter...such as research, collaboration across multiple disciplines, writing, project management, and program evaluation...process or the whole process, including grant readiness, funding opportunity research, application writing...

  12. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ... the speaker and offer your business card (yes, you should have a business card; it’s a bit more informal...smaller talks within the bigger conference, especially ones that are more career-focused. After these talks,... will probably be willing to accept your CV or business card. Be warned, though – someone accepting your... formally apply for the position. This post on Business Insider nicely summarizes good job fair practice... Reading and Outside Resources http://www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014...

  13. Bioinformatics at Addgene

    Type
    Blog Post
    ...engineering pipelines, we selected Nextflow, a workflow management system for scientific pipelines, including... existed who had created a shared resource of pipelines for genomic data. Thanks to the flexibility inherent...separate Nextflow process. In Nextflow, a process defines which command or script will run and can be thought... connect each subsequent process to the existing ones. Once our first process was working as expected,...check, we repeated our strategy and copied a few lines of our existing script into the section of the next...

  14. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...job it is to disagree with the group. They are the ones who ask for rationales behind ideas and decisions...fill out the form at any time and a member of our Business Development team would review it. This survey ...frequently observe in a scientific setting.  Powerlessness It’s hard to be creative when you don’t believe...ideas obviously worth pursuing, evaluate the “bad” ones. Bad ideas are easier to work with than nothing ... mistakes that kill your productivity. Harvard Business Review. Retrieved January 14, 2022, from https...exercise to spur innovation on your team. Harvard Business Review. Retrieved January 14, 2022, from https...

  15. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...love with biology because of an image that was honestly quite boring. My Bio 101 professor displayed a...mammalian vectors are even less similar to bacterial ones. The mammalian analogue of bacterial transformation...uptake, but this approach requires compatible cell lines that express viral machinery. Alternatively, some...selection is useful for producing stable mammalian cell lines, but it isn’t common for simple transfection experiments...by transformation and transfection and left the business of transduction to our Viral Vectors 101 series...Resources on Addgene.org Collection of empty plasmid backbones ...

  16. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ... transition Base change between purines (A ↔ G) or between pyrimidines (C ↔ T).   Base transversion ... ABE would allow it to act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the...this promising start, the team used rational mutagenesis to produce the adenine transversion editor AYBEv3...and ABEs, they could be useful for saturation mutagenesis, and they provide a strong proof of concept for...empty space. Over several rounds of rational mutagenesis, the team progressed to gGBEv6.3, which was capable...

  17. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...activators and repressors to target genes. Repressing target genes using dCas9-based repressors Early ...repression or knockdown allow you to study essential genes whose knockout would cause cell lethality. The Krüppel...purpose. dCas9-KRAB plasmids for repressing target genes in a variety of species and cell types can be found...found on Addgene’s website. Activating target genes using dCas9-based activators The simplest of the dCas9...transcriptional activator. Plasmids for activating target genes in a variety of species and cell types can be found...

  18. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...that recognize codons found in highly expressed genes (Emilsson and Kurland, 1990). Controlling gene expression...changing conditions - Organisms often need to express genes at different levels under different conditions. ...specific tRNA pools. For example, tRNAs used in genes encoding amino acid biosynthetic enzymes may be ... the problem of codon choice is to resynthesize genes in such a way that their codons are more appropriate...properly. You may be able to avoid getting your genes of interest codon optimized by ordering plasmids...use the same expression system for many different genes without having to create new constructs. However...

  19. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...in mouse embryos to generate gene modified mouse lines. Hopefully this post will help get your in vivo ...that later). Find Validated gRNAs to Target Mouse Genes To begin the in vitro screening process, you should... at the same time, and choose the most effective ones to increase your chances of gene modification in...a rapid acceleration in the production of mouse lines that could be used to study a wide variety of phenomena... et al. "Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes....generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering." Cell...

  20. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...Commercial Plasmid Prep Kit Amount 1000 Lanes 1000 Lanes 1000 Lanes Cost $200-$500  <$10 $20-30 Cost...commercial ladders cost between $200-$500 for 1000 lanes worth of ladder. The Penn State ladders are significantly...prep protocol, it cost less than $10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder. Using a ...
Showing: 321 - 340 of 902 results