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We narrowed to 729 results for: NSI;

Showing: 161 - 180 of 729 results

  1. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic...Rosella is a pH-sensitive fluorescent biosensor that was recently deposited with Addgene by Dr. Mark ... and recycling. The key to Rosella’s autophagy-sensing abilities is that its fluorescence emission spectra...regardless of its localization in the cell. SEP’s pH sensitivity means Rosella will fluoresce green unless it’...

  2. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...secondary considerations to validation, application specificity, and experimental considerations - no other...has been validated in your specific application. Consider also that there may be variations within an application... your needs.    Multiplex experiments Finally, consider whether you will be using other antibodies in ...from a vendor that you use frequently, with a responsive and knowledgeable technical support team, is ...

  3. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...spectral properties of the fluorophores you are considering to make sure that they’ll be compatible with ...antibody conjugates (especially if you start to consider clinical applications). But there are a few other... at least mention. Oligonucleotides allow for sensitive detection of low abundance targets with significant...mechanisms. Most take advantage of chemical groups intrinsically present on antibodies, such as the primary amine...can conjugate an antibody yourself allows you to consider a broader selection of antibodies for your experiments...

  4. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    Published
    Oct. 29, 2019, 2:52 p.m.
    ...channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization in continuous... the imbalance of electrical charge between the inside and the outside of the cell. This imbalance is ...Q239 - A GEVI based on the fusion of the voltage sensing domain of a Ciona intestinalis voltage sensor and...anion channel. It is triggered by a short low-intensity blue light pulse and stays open for a longer duration...

  5. Developing Lab Management Software for Biology

    Type
    Blog Post
    Published
    Aug. 25, 2016, 2:30 p.m.
    ... learned over the years with the hope that our insights can help steer you in the right direction when... of this is gospel, but we think it's worth consideration. Everything will go wrong Whenever an order ...easily as needed. In this case, the physical considerations of the laboratory make object composition a...technical details of the problem you're solving, but consistency is key. Some namespace collisions you may encounter...treating these pieces of data in ways that are inconsistent with how they are being treated in the lab, ...

  6. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    Published
    Feb. 11, 2014, 12:13 p.m.
    ...given problem, Empowers the writer, And offers insight into a career as a Professor. Step 1: Your ...sections in a NIH RFA, for example, and tips to consider while writing each section. Research challenge...time to review the literature and offer educated insight into what the desirable features of future technologies...deem your proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet... the application. Make sure to invest time and consider the reviewer's comments. Whether the proposal ...

  7. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...virion is a non-enveloped particle of ~100 nm, consisting of a capsid surrounding an inner core which contains...adenovirus: the virion is ~ 100nm in size, and consists of an icosahedral capsid made of 3 types of proteins... for creating adenoviral vector constructs. It consists of 2 plasmids: a shuttle vector (in which the ...purification of rAdV uses cesium chloride (CsCl) density gradients combined with ultracentrifugation to ... rAdV genomes/mL) can be measured by: Optical density: particle concentration is measured by looking ...

  8. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...Interactions Difficult to detect weak and/or transient PPIs. Many PPI identification techniques rely ...often prevents the identification of weak and/or transient interacting proteins. No spatiotemporal control...mass spec analysis. Biotinylated proteins are considered possible interaction partners if they are enriched...BioID, is also useful for identifying weak and/or transient protein-protein interactions since the biotin ...cell lysis and lysate fractionation. Important considerations for Split-BioID It’s important to remember ...

  9. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles. Depending on the researcher...ultracentrifugation of the viral stock, followed by resuspension of the collected particles in a smaller volume...Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and should be stored in ...enhance transduction efficiency in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic...PMID: 12050007. 7. Stockschläder, M., et al. "Expansion and fibronectin-enhanced retroviral transduction...

  10. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ribosomal... Rho-independent termination is also known as intrinsic termination, and it relies on the formation of...ribosomal RNA, Polymerase II is responsible for mRNA and miRNAs, and Polymerase III transcribes tRNA and other...polymerase-associated proteins, CPSF and CstF, which are responsible for recruiting the cleavage and polyadenylation...

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

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...because they are inside the virus itself. While these impurities are generally considered innocuous for ...helper plasmids or the cell genome can be packaged inside the vectors during production. These impurities...NGS at Addgene to sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged...contamination is a good start, but to determine the sensitivity of our method we wanted to see how low we could...

  12. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...this temperature and can bind to the primers.  Extension- The temperature is then raised slightly to Taq...  This process of denaturation, annealing and extension is repeated 25-35 times to exponentially replicate...droplet PCR or ddPCR is a method that provides ultrasensitive and absolute nucleotide concentration, unlike...more complicated to develop and is often less sensitive than PCRs that use a single pair of primers, like...Diagnostics NGS library prep QC ddPCR Determine ultrasensitive and absolute nucleotide concentration of starting...

  13. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...injections are most often used when injecting a cell suspension to create xenograft models. Restraint: back scruffing...sounds very disruptive, but surprisingly, it is considered minimally invasive. Restraint: anesthesia Location...common. Due to the intubation, this procedure is considered quite invasive. Restraint: anesthesia Location...introduction to common injection routes in mice! When considering what method you’d like to use, remember that ...animals: routes of administration and factors to consider. PubMed, 50(5), 600–613. https://pubmed.ncbi.nlm.nih.gov...

  14. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...to their high density, which can be collected afterwards. You can then remove the density gradient material...certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin...cells and supernatant then need to be purified by density gradient centrifugation (e.g., iodixanol) using...assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events...

  15. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    Published
    Dec. 12, 2013, 3:35 p.m.
    ...that onto a mass spec. That's very time intensive and expensive. There's the promise that we could instead...DNA systems capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks...describing the development of “transcriptors,” transistor-like digital genetic switches that enable cellular...proteins. That's central to everything that goes on inside a cell. Until we get a handle on precise, forward...

  16. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN is the N-terminal...(Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion proteins N713-pMag and nMagHigh1...able to reach deeper tissues. Additionally, high intensity exposure can cause DNA damage. Far-red light, ...Michael Lin, who developed a chimeric protein consisting of Cas9 and a photo-dissociable dimeric green... anti-CRISPR Acr protein, AcrIIA4, to a photo-sensitive LOV2 domain from A. sativa (Bubeck et al., 2018...editing For more details, check out this comprehensive review comparing optogenetic CRISPR techniques...

  17. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...are some potential complications that must be considered when using this method:     Cells must be amenable...to directly deliver a ribonucleoprotein (RNP), consisting of the Cas9 protein in complex with a targeting...gRNA from a commercial vendor, this can often be expensive. An alternative approach is to express and purify...is most appropriate for a given experiment. Considerations for RNP-mediated CRISPR experiments While using...with these types of experiments. One important consideration is that the Cas9-gRNA RNP will be degraded by...

  18. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    Published
    Jan. 11, 2018, 3:29 p.m.
    ... venture presents itself even outside of work, consider challenging yourself, take the opportunity, and... all the things for which you are officially responsible on paper, and take some time to write down all... have to exercise in order to manage these responsibilities. If you are in charge of completing many ...management and organizational skills. If you are responsible for working with lots of students, managing inventories...the short list of transferrable skills here and consider which of these skills you’ve used in your own ...

  19. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...immune system directly). It may seem odder when you consider that out of the 6 x 106 virus-like particles that...DNA called a “provirus.” This DNA is synthesized inside of what is called the “target cell” (or host cell...reverse transcriptase are packaged (Wu et al., 1997) inside of the capsid . This is useful for viral reproduction... particle, some of the RT protein gets packaged inside of the vector particle, along with its RT-deficient... natural virus, this new chromosomal locus is considered “proviral DNA” (Rowe et al., 1971) and the full...

  20. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...use a standard curve and provide some general considerations for scientists planning to use them in their...to 1:32. Each individual step of the series is consistent at 1:2. When preparing a dilution series, use...users aim for an R2 above 0.95.    Additional considerations When running assays that rely on a standard...not fall within this range, it may be time to reconsider your graphing program. Summary Few things in ...run.  When setting up your dilution series, use consistent dilution factors, change pipette tips regularly...
Showing: 161 - 180 of 729 results