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We narrowed to 727 results for: cat.1

Showing: 421 - 450 of 727 results

  1. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...CRISPR tools.  Distributing and depositing In Figure 1, you can see cumulative numbers of CRISPR plasmid ...new CRISPR depositors each year.    Figure 1: Addgene CRISPR distribution and deposits. (A) Cumulative...estimated scatterplot smoothing. (C) Number of requests per year for pooled libraries, categorized by function...insertion via a chemical modifier. We used these categories to see what kinds of CRISPR tools people were... pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2, the numbers...Cumulative number of deposited CRISPR items, categorized by CRISPR function, by year. (B) Number of requests... years.  Libraries are divided into different categories based on function: knockout (KO), activate, inhibit...

  2. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...interrogated by the optical system (Figure 1).   Figure 1: Cells expressing a particular surface ...populations of cells in a complex tissue or sample (1). When combined with cell sorters, this technology... it passes through and scatters in different directions. The degree of scatter is captured by detectors...Light that scatters sideways to the path that it was initially traveling is called side scatter (SSC) and...granulocytes in a plot of forward scatter (FSC) versus side scatter (SSC). Users can gate around their...protein composition. It is used for a variety of applications such as measuring target protein expression ...expression levels, assessing post-translational modifications, determining cell health, analyzing cell cycle stages...

  3. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes... dopamine D2/D3 receptors. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-78827-9 Breen... Biophysica Acta - Biomembranes (Vol. 1838, Issue 1 PARTA, pp. 15–33). https://doi.org/10.1016/j.bbamem...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...whereby excitation of blue light will indicate where FITC is bound through green light emission. Key considerations...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...

  4. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...found climbing mountains or playing with his two cats. Keira Havens grew up in Hawaii where she was fascinated...challenges involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the...biotechnologists face is public perception of genetic modification (GM) and synthetic biology. The mistrust and...cannot build what we do not understand, and the complicated and unique metabolic processes of plants are ...

  5. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes...dCas to the genomic DNA of interest  Figure 1. When ABEs are in action they bind DNA so that the... from E.coli, named TadA7.10 in ABE7.10)  a catalytically impaired Cas protein (dCas or Cas nickase)  ...accidental insertions and deletions (indel) by the catalytically impaired D10A nickase from ABE7.10 can be a ...the authors created ABE8 constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The...these variants, ABE8 could target many additional locations in the genome. Compared to ABE7.10, NG-ABE8.xm... reduced off-target editing frequencies. For applications that require high DNA editing specificity, for...

  6. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...fluorophores are constantly being engineered. Table 1 lists a few common fluorescent proteins and their ...definitions found in Table 2 may not apply. Table 1: The specific mutations comprising common fluorophores...transgenic mice. Purification: GFP can be used as a general epitope tag for protein purification and a number...their experimental needs, and some of the common applications in the lab. Why Green Fluorescent Protein?...to maintain fluorescent activity; very little truncation is tolerated, however, point mutations are acceptable...Re-engineering GFP to increase its range of color and application Almost as soon as its sequence was elucidated...others) Neutral mutations    A multitude of applications Due to its size and ease of use, GFP and other...

  7. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can block CRISPR activity... ways to anti-CRISPR proteins in your experiments 1. Reduce off-target effects Prolonged Cas activity ..., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered that...viral particles that have the gene deletion could replicate when challenged with the S. islandicus CRISPR-...B, Bondy-Denomy J (2020) Anti-CRISPR protein applications: natural brakes for CRISPR-Cas technologies....light-inducible Cas9 activity? Check out these these plasmids Catch Up on Your CRISPR Background with Our Guide Pages...

  8. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...crevices proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody...Humana Press, pp 133–148. https://doi.org/10.1007/978-1-62703-293-3_10 Walter J et al. (2020) Synthetic nanobodies...Starting with libraries and a biotinylated (for purification purposes) protein target of interest, the protocol...-30) of sybody candidates are sequenced for identification. All unique sybodies can be subcloned into ...2020) Generation of synthetic nanobodies against delicate proteins. Nature Protocols 15:1707–1741. https...Light to Manipulate Cell Signaling and Protein Purification RANbodies: Reporter Nanobody Fusions Plasmids...

  9. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...Updated Jun 1, 2021 by Meghan Rego. Reproducible data are key to science, so scientists are used to repeating...the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from purified...These impurities cannot be removed during the purification step because they are inside the virus itself...very low concentration. So how do we make sure to catch all of these potential mishaps before distributing...Guerin et al., 2020).  In addition to genome identification, we developed custom open-access Python scripts...

  10. Of Myc and Men

    Type
    Blog Post
    ...used in way more applications.   Fig. 1 Making a Protein Tag (Abridged) (1) Locate a promising monoclonal... by its antibody without additional chemical modification steps and when the epitope was within the tagged...

  12. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...for example, split cells on a Monday diluting them 1:10, you should be able to split on the following Thursday...Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...Anything between orange and light orange usually indicates that the cells have grown enough, are making their...

  13. Plasmids 101: Protein tags

    Type
    Blog Post
    ...DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based purification; has inherent enterokinase...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function...site GST Large Protein 26 Purification and Stability Good for purification with glutathione; protects...used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity and Purification Very small...GKPIPNPLLGLDST   1.4 Affinity and Purification Good for antibody-based purification Combo and cleavage tags...multitude of uses including (but not limited to) purification, detection, solubilization, localization, or...fusion proteins to aid in the expression and purification of recombinant proteins. Tags for stability...

  14. Hot Plasmids - November 2023

    Type
    Blog Post
    ... edit.   Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman...delivery by modular envelope design. Nature Communications, 14(1), 5141. https://doi.org/10.1038/s41467-023... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting... (which had therefore been trafficked between locations). Figure 3: A) LOV-Turbo becomes active...active when illuminated with blue light, catalyzing the biotinylation of nearby proteins. B) TransitID uses...

  15. Hot Plasmids - August 2020

    Type
    Blog Post
    ...membrane targetin sequence derived either from p63(1-29) and Lck. Image from Chertkova et al., 2020. ...Reports. 2020. https://doi.org/10.1038/s41598-020-57500-1 The CRISPR corner By Jennifer Tsang Here are a few...Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system for hPSCs The CRISPR...CRISPR corner New from the viral service Faster catalysis after yeast-based protein evolution By Joanne ...concept, Sanchez and Ting sought to improve the catalytic efficiency of tobacco etch virus protease (TEV...activity with a fluorescent protein--no affinity purification required! The team used other molecular techniques...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor ...

  16. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631. PubMed PMID: 22368488...be selected for these small ligands. References 1. Ma, Hanhui, et al. “Multicolor CRISPR labeling of... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors... render this approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...

  17. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...gamma-retroviruses ranges from 9–11 kb, encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus...producing viral vectors in a laboratory setting. Figure 1: Wild-type gamma-retrovirus genome. Created with BioRender.com...and tropism. Replication-competent The ability of a virus (or viral vector) to replicate and reproduce...hosts in order to continue their lifecycle and replicate. This feature can be harnessed for stable delivery... for safety to reduce the chance of creating replication-competent retroviruses. For a summary of all ... gamma-retroviral vectors integrate at random locations throughout the genome. Gamma-retroviral vectors...retroviral vectors are commonly used in gene therapy applications to more specifically direct delivery of gene...

  18. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...Spaghetti Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins... expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes...controlled by collections of neurons that are intricately wired into circuits through synaptic connections...photostable than GFP. In addition, antibody-based amplification of their signals is weak and increases background...more HA tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional microscopes...

  19. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...regulated in the medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our research ...activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong...and a vacuum is applied with a vacuum bell or desiccator. Successful infiltration can often be seen by...expression method for analyzing gene function in Catharanthus roseus seedlings." Frontiers in plant science...

  20. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ... cerevisiae (Tuller et. al., 2010). Figure 1: An overview of peptide synthesis. A ribosome interacts...McClain WH (2000) The G·U wobble base pair. EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001...folding and stability, change post-translational modification sites and even affect protein function (Mauro... AP, Bilen B, Zavolan M (2013) Analysis of CDS-located miRNA target sites suggests that they can effectively...synonymous codon usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org...

  21. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...about health equity, science communication, and public health.     References 1. O'Neill, J. "Tackling drug-resistant... to the development of resistance include (Figure 1): Over-prescription of antibiotics (for example, ...Microbes (bacteria included) are very skilled at the delicate art of survival. Existing virtually ubiquitously...of intentions. The take-home? Always take your medication as prescribed. How do we know which bacteria ...

  22. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ... present in the sample (APC in the case of Figure 1). In Figure 1A, the population needs to be shifted... decrease the compensation value.   Figure 1: Compensation example with single color control. Flow...diagonal population, this is a sign of spillover and indicates that it is necessary to apply a compensation value...

  23. Countdown to Halloween @Addgene

    Type
    Blog Post
    ...glorious trophy. Sure it’s plastic and you have to share 1 trophy among 6 people, but you’ve earned bragging ...plasmids! Each chose their favorite plasmid and replicated it. Can you guess each plasmid? Our lab paid...creative hijinks last year. They showed us their dedication to great customer support by dressing up as our...

  24. New Viral Vectors - Spring 2025

    Type
    Blog Post
    ... Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor...skipping the preparation steps of production, purification, and titering. Our newest viral vector service...

  25. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    ...a ‘plasmid’. “p” is for plasmid. pXXXXX-XXXX Step 1: Backbone name Include the empty backbone name in ... eukaryotic cells). pBACKBONE-XXXXX Tip: For a catalog of published and commercially available empty backbones...considerations  If your insert contains a mutation or modification, this should be included in the plasmid name...

  27. Streaking for Single Colonies: The Streak Plate Challenge

    Type
    Blog Post
    ...Time for the #BioSci3319 #StreakPlateChallenge part 1. Lab B needs a redo :-( @SandTBioSci #CompetitiveStreaking...blog posts on fluorescent proteins Check out educational resources on the blog Resources on Addgene.org...

  28. New Viral Vectors - Summer 2024

    Type
    Blog Post
    ... Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor...

  29. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ... necessary for advancing the field. References: 1. Internal initiation of translation of eukaryotic ... of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. Kim et al...desktop. The Plasmids 101 eBook is designed to educate all levels of scientists and plasmid lovers and...

  30. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...true reproducibility. The following are critical: (1) connectivity to other data sources used in research...engagement with the Wisconsin research community were to: (1) prevent vendor lock in by making it easy to get data... both accept deposit of data associated with publications into one of five general purpose open source...
Showing: 421 - 450 of 727 results