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We narrowed to 634 results for: des.2

Showing: 341 - 360 of 634 results

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

    Type
    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...biochemical properties. In 1995, Roger Y. Tsien described an S65T point mutation that increased the fluorescence... in the wildtype protein and facilitating its widespread use in research. Many other mutations have since...array of GFP-based fluorescent biosensors has been designed to detect a variety of intracellular conditions...

  2. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody... and to work in a variety of applications. The desired specificity of your secondary antibody depends ...

  3. 6 Tips for Grant Writing

    Type
    Blog Post
    ...days before the deadline isn’t going to be much use. 2. Originality is essential for grant success The two... it’s important not to throw in the towel if 1 or 2 funding bodies say no. If possible, get feedback from...to a lay audience. My lay paragraph typically includes a couple of sentences of introduction to my area...

  4. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...improve antibody binding for some targets, they can destroy it for others. That said, organic solvents’ impact...critical to the outcome of your IF experiment and deserve careful thought and attention.   * There are other...

  5. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression...Patrick Miller-Rhodes from the University of Rochester Medical Center. Patrick Miller-Rhodes is currently...This post was contributed by Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at ...this technological gap, a trio of recent papers describe the development of CRISPR activation (CRISPRa)... large size of Cas9 activators, which largely precludes delivery by adeno-associated vectors (AAV). This...packaging capacity than AAV. Such an approach was described recently by the Day Lab, who developed neuron-...

  6. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ...reproducibility than others. Unfortunately, most ELNs are designed as closed ecosystems and act as data silos because...RSpace ELN supports reproducibility RSpace was designed in response to requirements that emerged from ...used in research, and data repositories. Below I describe how each of these kinds of connectivity facilitates... in institutional and lab servers specifically designed to hold large amounts of data. Although it may... use of other tools they use in research. This includes chat apps like Slack, Microsoft Teams, and Google...

  7. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ... which one you choose depends on your experiment (de Ruijter et al., 2003). For example, we did our initial...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...Humana Press, 2009. 41-50. PubMed PMID: 19183892. De Ruijter, N. C. A., et al. "Evaluation and comparison...and a vacuum is applied with a vacuum bell or desiccator. Successful infiltration can often be seen by...

  8. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    ...important role in solving the structure of DNA. 2. The Immortal Rap of Henrietta Lacks Students in Oakland...One of Tom's earliest and most popular videos. Describes fossil rocks and their importance in understanding...

  9. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...human therapeutics (e.g. using Sleeping Beauty). Despite their diverse utility, DNA transposons have been...(V. cholerae INTEGRATE - hereafter VchINT) and encodes a Type I-F CRISPR-Cas system. The four major components...the same promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also...: Clone spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid...even very large genetic cargo, at one or multiple desired genomic loci. Many applications of these systems...

  10. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...individual molecules. Poor photon budgets are a widespread issue with fluorescent proteins—even the thriftiest...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical... the complexity of the system. A particularly desirable property for small-molecule labeling strategies...size, brightness, and cell permeability. In past decades, further refinements to this established dye structure...spectral range. Nevertheless, these fluorophores were designed as antibody and oligonucleotide labels for fixed-cell...

  11. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...Over the past decades, neuroscientists have made great strides to map brain regions and allocate specific...for this task. However, most lesioning methods destroy areas larger than intended and may unintentionally...surface. The latter reflects a state of behavioral despair, and the more time the animal spends in the passive...that were not (Hare et al., 2019). The research described here is only a small selection of the increasing...

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

    Type
    Blog Post
    ...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial... defined as microbial survival despite exposure to antibiotics designed to kill them or to impede their...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...different reasons. Exposure to antibacterial drugs provides a selection pressure, killing off bacteria that...equipment, etc. Virtually any preventive measure designed to help keep you from getting sick can be helpful...

  13. Countdown to Halloween @Addgene

    Type
    Blog Post
    ...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween...for Halloween and how seriously we take costume design. In case you need more convincing, I’ve got just...to great customer support by dressing up as our Zendesk email system.  Our 2013 winners dressed up as ...

  14. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I ...The NPTII gene confers resistance to several aminoglycosides, including G418 which is commonly used for ...DNA and generating DNA breaks. The Sh ble gene provides zeocin resistance by expressing a protein that...experiments in which off-target mutations are undesirable. Pros: Works across many species/systems. Cons...

  15. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ...Microscopy. What is Expansion Microscopy?  ExM provides a straightforward answer to the space problem....improve label density (Mikhaylova, et al. 2015).  Despite these limitations, ExM is a valuable tool, especially...Bournonville L, et al (2021) Expansion microscopy provides new insights into the cytoskeleton of malaria ...

  16. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...most situations.  A slightly different method is desirable if the antibody is expected to bind weakly to ...vary in shape and structure. Their porous surface provides a large area for binding, but antibodies conjugated... slightly higher, than those of agarose beads, despite the size difference. Magnetic beads are particularly...

  17. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ... editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with...follow-up posts: (1) (2). Base editors have a narrow editing window of only a few nucleotides, so they are tightly... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...

  18. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....case, a buffer exchange can be performed using a desalting column or an ultrafiltration concentrator. As ...suggests, you may also concentrate your prep to the desired concentration using these columns. If you are conjugating...

  19. To Each HIS Own

    Type
    Blog Post
    ...take care when using this method.      Fig 2:  A, The polyhistidine tag is a string of histidine... Bio/Technology. 1988; 6: 1321–1325. Janknecht R, de Martynoff G, Lou J, Hipskind RA, Nordheim A, Stunnenberg... for nonspecific binding of untagged proteins. Despite being a relatively low frequency amino acid, proteins... I place the tag and how many do I need? When designing a plasmid containing a polyhistidine tag, you’...

  20. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity for in vivo studies...single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation...assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission...also reveal novel pathways. The FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction...
Showing: 341 - 360 of 634 results