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We narrowed to 672 results for: abo.1

Showing: 121 - 140 of 672 results

  1. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ... microscopy contrast technique in modern biology (1). When performed correctly, fluorescence microscopy...relatively thicker three dimensional sample (see Figure 1); similar to an MRI or CT scanner. This is accomplished... the resolution of a light microscope (see Figure 1). What microscopy technique should I use for my experiment...new sample I always start by asking two questions: 1) Is the sample dynamic or static? 2) Is the sample...of modern day fluorescence microscopes (see Table 1). Thin dynamic samples Ex: monolayers of live cells...excitation light can push imaging depths closer to 1 mm. Traditionally, 2-photon has been a very slow imaging...manufacturers have similar information References 1. Lichtman JW, Conchello JA. Fluorescence microscopy...

  2. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ...not let the panelists introduce themselves. I do a 1-2 minute introduction. If you chose good panelists...take 5 minutes to introduce themselves and, voila, 1/2 of your time is gone with no interesting discussion...introductions. The moderator asks questions, calling on 1-2 panelist for each question in a way that elicits... time you are at a talk and the speaker goes over 1 hour, watch what happens to the attention of the audience...postures shift, they are checking out. If you can, plant 1-2 good questions with trusted audience members. A ... are nice if you can afford them). I am talking about making sure the audience is interested, educated...of the stories or topics you know they can speak about. There should be some spontaneity, but having a ...

  3. Antibodies 101: Producing Recombinant Antibodies

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    Blog Post
    ...materials have been washed away (Figure 1).      Figure 1: The steps of rAb production. Created ...EBNA1 Cells. Methods Mol Biol. 2018;1850:1-16. doi: 10.1007/978-1-4939-8730-6_1. PMID: 30242676. Frenzel...Neuroscience Research. Curr Protoc Neurosci. 2020 Dec;94(1):e106. doi: 10.1002/cpns.106. PMID: 33151027; PMCID...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...validated, it’s ready for use! You can learn more about common antibody applications in our Antibodies 101...

  4. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...protocols. A typical ExM protocol (Fig. 1) follows this structure: Step 1: fix (and maybe label) your sample...microscopy. Nat Biotechnol 1–12. https://doi.org/10.1038/s41587-022-01546-1  Mikhaylova M, Cloin BMC, ...just adding water.)  Step 5: Image   Figure 1: The ExM process allows you to physically magnify ...16:71–74. https://doi.org/10.1038/s41592-018-0238-1  Klimas A, Gallagher BR, Wijesekara P, et al (2023...applications and there’s just not much to be done about that.  Second, expansion introduces distortions....

  5. Negative Can Be Positive: Open AAV Data with Addgene

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    Blog Post
    ...across scientific disciplines. Scientific Data, 8(1), 1–11. https://doi.org/10.1038/s41597-021-00981-0 Wright...excellent at infecting our target neurons (Figure 1). However, this one set of experiments didn’t provide...yet none of that data was published.    Fig. 1: Testing of three different AAV serotypes (AAV1, 8...purification and formulation. Molecular Therapy, 12(1), 171–178. https://doi.org/10.1016/j.ymthe.2005.02.021...Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https://doi.org/10.3389/fnhum.2020.578687 Additional...asked why they don’t share, many express concern about sharing negative data they cannot explain, perhaps...

  6. 500,000 Plasmids Shipped and Counting!

    Type
    Blog Post
    ...here at Addgene – 500,000 plasmids shipped! That's a 1/2 million plasmids sent to happy scientists around...start out at our Cambridge office – previously at 1 Kendall Square, but newly relocated to 75 Sidney Street...Have a question? Email: [email protected] Call: +1 (617) 225-9000         ...scientists outside of the US now use the repository – about 50% of plasmids are shipped to labs outside of the...distribute their plasmids. The partnerships and collaborations with institutions and tech transfer offices...

  7. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...Correct Path.,” J. stem cell Regen. Biol., vol. 1, no. 1, Jan. PubMed PMID: 26702451. PubMed Central PMCID...genetic manipulation in mammalian cells until 2005 [1]. Like other transposons, piggyBac™ has two components...The predicted frequency of ‘TTAA’ in the genome is 1 in every 256 base-pairs of DNA sequence, making it...has been demonstrated) with no known upper limit [1]. The piggyBac™ technology can be used for numerous...genetic loci of integration [8].   Figure 1: Mechanism of the piggyBac™  transposase/transposon...technologies and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-ing models...transposase itself, please read on to learn more about this exciting technology from the folks at Transposagen...

  8. AAV Packaged on Request is Here!

    Type
    Blog Post
    ...volume, and submit your request.    Figure 1: The AAV Packaged on Request banner.   We currently..., AAV8, or AAV Retrograde (AAVrg), and volumes of 1 mL, 2.5 mL, or 5 mL. Please note that all orders will... our on-demand viral service, we expect titers of 1 × 1013 GC/mL for AAV1, AAV5, AAV8, and AAVrg and 7...your browsing and requesting. If you’re curious about the service, read on.  Expanding our viral vector...

  9. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...building it! Table 1 has a list of Fragmid destination vectors currently available. Table 1: Fragmid destination...components on the Fragmid website.   Figure 1: (Top) Schematic overview and timeline of the Golden... CRISPR vector the Fragmid way!    Figure 1: Selecting your vector type on the Fragmid website...components available will depend on your answers in step 1, so you’ll only be able to select components that ...Destination vector (dropdown options depending on above selections) Module set (the set of components you...vectors) Target type (drop down options depending on above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other...

  10. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...also important for social and regulatory reasons. 1.) The transposon consists of a mirrored set of inverted...vertebrates. The system is comprised of 2 components: 1) an SB transposase, the enzyme required for catalysis...likely to grow in the coming decades.  References 1. Kowarz E, et al. Optimized Sleeping Beauty transposons...T cells to target CD19. Immunol Rev. 2014 Jan;257(1):181-90. doi: 10.1111/imr.12137. PubMed. Find the ...of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both introduce...

  11. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...including yourself as the researcher.   Figure 1: Separation of retroviral packaging components onto...these reasons, AAV is usually classified as a BSL-1 safety level. Adenovirus is similar to AAV in that...lab coats are either recommended or required in BSL-1 and required in BSL-2 labs. When working with lentivirus...required whenever there’s a splash or spray risk in BSL-1. In BSL-2 labs, eye protection is required any time...isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some organizations or cities may...completely inert, but others can cause diseases. Nervous about handling viruses? That’s okay! In this article we...

  12. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...type of assay is a clonal cell line known to contain 1 integrated copy of the qPCR target gene. Developing... successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob Reiser...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009... titration methods."BMC biotechnology 6.1 (2006): 1. PubMed PMID: 16836756. PubMed Central PMCID: PMC1534021...events." Genetic vaccines and therapy 2.1 (2004): 1. PubMed PMID: 15291957. PubMed Central PMCID: PMC514534...drawback of this approach is that it can be quite labor intensive. Target cells are transduced with serial...

  13. Technical Design of a Western Blot

    Type
    Blog Post
    ...Tris-tricine 2.5–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours Good separation, quality, and stability...lot to cover, so let's dive in!      Figure 1: The technical decisions to make when designing a ...assay. The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose...overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce...weight of the proteins you’re interested in. Table 1 lists the most common types of gels for SDS-PAGE running...gel chemistry and/or specialty gel options. Table 1: Types of gels  Gel Type Protein sizes Running... Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life...

  14. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...the genome, which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and NgAgo genome engineering...it was first published in Nat Biotech in May 2016 (1), NgAgo has received much attention and the plasmid...Pooran's NgAgo Survey Survey results As of August 1, 2016, a total of 165 researchers responded to the...the cell, the same method used by Gao et al, 2016 (1). View the Complete and Up-to-Date Results of Pooran's... dreams of a sunny day in Edinburgh.   References 1. Gao, Feng, et al. "DNA-guided genome editing using...technology. To dig more into what researchers felt about NgAgo, I conducted a survey and asked researchers...successful, and the second section asked basic questions about the NgAgo protocols used by respondents (transfection...

  15. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...choosing direction of a project. If you expect more 1:1 time with the advisor, you should choose accordingly...hire visits the lab for a full day, gives a 1 hour talk about his or her interests, and spends one-on-one...that supports both paths – you won't know in year 1 what you will want in year 6. Have lab alumni taken...during the day? We asked three lab heads how they go about selecting new hires. This is followed by some perhaps...current members to share short-term goals and chat about awesome science. Important factors in the search..., the lab has a reputation for not entering collaborations. Alternatively, perhaps the lab publishes and...Medical School) for taking the time to speak with us about adding new scientists to their labs. Read the Other...

  16. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    ...animal, those B cells are fused to myeloma cells at a 1:1 ratio and then cultured in a special medium called...antibodies that recognize proteins. Figure 1: Steps in the generation of hybridomas for monoclonal...Washington (DC): National Academies Press (US); 1999. 1, Generation of Hybridomas: Permanent Cell Lines Secreting... Learn more about research applications for antibodies in Intro to Antibodies Read about the Developmental...antibodies in your experiments, you may be curious about all of the different kinds of antibodies that are...Disadvantages of monoclonal antibodies As mentioned above, genetic drift is one concern when using hybridomas...Hybridoma Bank, a repository for hybridomas  Learn about monoclonal antibodies for immunotherapy Resource...

  17. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...cellular proteins are modified through phosphorylation (1). However, the expansion of studies on protein phosphorylation...three components required for translation (Figure 1): A tRNA that recognizes a specific ‘open’ codon ... open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...modification allowed the deletion of release factor 1, the protein responsible for terminating translation...Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing...follow her on Twitter @nieminm.   References 1. Cohen, Philip. "The origins of protein phosphorylation." Nature... one of the MEK1 phosphomimetic residues in the above study with phosphoserine (to create a pSer/phosphomimetic...

  18. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

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    Blog Post
    ...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...which have previously been characterized. See Box 1 in Kudo et al. for more details on the design of the...using translocation reporters. Nature protocols, 13 1, 155-169. PMID: 29266096. Find the KTR plasmids here...Blog Learn about photoswitchable kinases Learn more about fluorescent biosensors Learn about using phosphoserine...

  19. Plasmids 101: Secondary Nanobody Toolbox

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    Blog Post
    ...incubation of 1° and 2° Must use different species and/or different IgG subclasses of 1° for multi-color...displacement 2° incubation can be skipped by pre-binding to 1° Monovalent Monoclonal Only anti- rabbit and -...dyes are also commercially available. References 1. Pleiner, T., Bates, M., & Görlich, D. (2017). A toolbox...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...Control Protein Activity Learn more about protein tags Read about E. coli strains for protein expression..., if not better, than antibodies for all of the above listed molecular techniques, but they can also be...protein purification methods. Read on to learn more about nanobodies and how their structure and function ...

  20. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...two-step, 2 hr protocol prior to transformation (Figure 1). As this approach requires no special reagents or...with linearized vector backbone. DpnI digestion - 1 μl DpnI is added to the PCR mix to remove parental...confirm primer design.  Use minimal template DNA Use 1 ng of PCR template DNA to minimize false positive ... by PCR, so we usually add DMSO (3%) and Betaine (1 M) to the PCR mix. When PCR is not possible Some plasmids...Twitter @jakefwatson and @JGarciaNafria.  References 1. Jones, Douglas H., and Bruce H. Howard. "A rapid ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...
Showing: 121 - 140 of 672 results