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We narrowed to 876 results for: NSI;

Showing: 161 - 180 of 876 results

  1. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...develops an enhanced propensity for movement and activity called locomotor sensitization. This enhanced movement... modulating. Viral vectors, in contrast, are considerably more versatile and specific in how they manipulate...ve previously discussed some of the general considerations for performing behavioral tests, but now we...seeking drugs’).  What’s involved in locomotor sensitization? Previous research has shown that regions such... glutamate transmission leads to locomotion sensitization. (for review see van Huijstee & Mansvelder  ...suggested inhibitory mechanism of locomotor sensitization and might open new ways for a novel therapeutic...subset of excitatory neurons (Drd1) in mPFC is responsible for the antidepressant effect. To specifically...

  2. Lab to Office Culture Shock

    Type
    Blog Post
    ...come up with a list of things to consider as you make this transition. Of course, a “desk job” could entail...employee who's coming directly from grad school. The transition has been a bit rough for her (3 weeks in and ...there are any resources that can help with the transition.” The lab-to-desk-job culture shock is a real...scientist colleagues have as well. Making the transition from the bench can be jarring for the unprepared...resources out there for scientists making this transition. In this installment of the Addgene career guide...What should you expect during the lab → office transition? I’ve already provided a few counterpoints to...likely happen eventually. If you approach that transition dreading the change, the adjustment period will...

  3. Working 9-5: an Addgene Co-op

    Type
    Blog Post
    ...training schedule that helped me transition easily. Training and responsibilities Through this training, I was...The application process When I first saw the comprehensive job description for this co-op, I was immediately...me a full description of the tasks I would be responsible for, in comparison to the other positions listed...for the independent work on the QC team. I was responsible for helping other scientists on the team ensure...sequencing QC work    In addition to my responsibilities on the QC team, I was able to explore other...uses something called “core hours”. Core hours consist of the time between 10 AM and 3 PM, when most meetings... the “9-5” was not as scary as it seemed. The transition into a traditional work environment was aided...

  4. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

    Type
    Blog Post
    ...Optogenetics, the use of light sensitive proteins (opsins) to manipulate cell activity, enables researchers... with fluorescent tags and activation of light sensing proteins. Furthermore, during the first 2 weeks...Optogenetics functions through opsins which are light sensitive proteins. Opsins used in optogenetic studies are...these charged particles and whether they make the inside of the neuron more positive or more negative.  ...  Neuronal stimulation: Depolarization When the inside of the neuron becomes positive over a certain threshold...inhibition: Hyperpolarization Conversely, when the inside of the neuron becomes more negative it becomes ...differing peak absorption wavelengths and light intensity requirements of different opsins, studies have...

  5. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ... being considered for the analysis. (C) A more liberal gate allows more cells to be considered for the... SSC values depend on the structural complexity inside the cell or on its surface. Bear in mind, the voltage...histogram, which graphically represents the time and intensity of the cell’s illumination (see Figure 2). H describes...potential that can be applied to the photomultiplier inside the cytometer to increase the electric current ...as our cells of interest do not lie beyond the dimensions displayed in the plot. (B) A more restrictive... marker stains of interest). Let’s double-click inside our gate and proceed to the next plot! Pro tip!... on to my experimentally-specific gates. Two-dimensional staining (quadrant gating) For our last example...

  6. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...A major limitation of APEX was its low sensitivity. Sensitivity could be increased with higher expression... avidin, proximity labelling can capture weak/transient protein interactions as well as be used for both...promiscuous biotin ligase (Kim et al 2016). BioID2 consists of a humanized biotin ligase from A. aeolicus,...cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate peroxidase for proximity labeling A ...aggregation. The Ting lab hypothesized that the low sensitivity was due to suboptimal folding or stability. Thus...identified a mutant with significantly increased sensitivity that did not affect the function or efficiency... assemble when in close proximity. Split-BioID consists of a split BirA* molecule that can reassemble ...

  7. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ... same considerations apply as for standard IF. However, there are a few specific considerations to keep...standard immunofluorescence, but with some extra considerations (Im et al., 2019; McLaughlin, 2019).    ...a good place to get started.  One important consideration when deciding whether you can do simultaneous...suitable antibody against your target protein, consider fusing a protein tag to the target and using an...epitope-based strategy such as Anti-6xHis. You could also consider strategies using other types of affinity reagents...your experiment, you might need to quantify the intensity of the different signals in a certain region or...optimizing your blocking and washing conditions or consider using a different antibody.  Prepare a positive...

  8. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...postsynaptic receptors and transporters. The intensity-based Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed...this post: Glutamate indicators with increased sensitivity and tailored deactivation rates Addgene's own...interneurons Glutamate indicators with increased sensitivity and tailored deactivation rates By Mike LacyGlutamate...significantly improved brightness, kinetics, and sensitivity. iGluSnFR4f is well-suited for imaging rapid ... (2025). Glutamate indicators with increased sensitivity and tailored deactivation rates. bioRxiv, 2025.03.20.643984...vector that contains everything needed to replicate inside a host cell but with space to insert your genetic..., GPC3, and GPC4). All IPI antibodies undergo extensive characterization at IPI as well as community-led...

  9. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ... are considering a career at a small, teaching college you might want to consider less expensive, portable...low salary I might add). You have a right and responsibility to choose a lab where you can thrive and do...Former lab members can give you a fuller picture. Consider talking to people in the lab across the hall or... hope to grow.  Find mentors who take their responsibility to train you seriously. Why work for someone...whether or not a lab head is a good role model, consider questions like: How long do lab members stay ...allocation of meaningless tasks, removal of responsibility, repeated reminders of blunders, setting target...equipment quotes? Understand administrative responsibilities? Is the work (topic and/or technical aspects...

  10. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...variation between iPSC clones. Retroviruses are not considered safe for clinical applications.     Lentiviral...Like retroviruses, lentiviral delivery is not considered safe for clinical applications. Excisable Cre-lox...same as using a standard lentiviral vector, but transient expression of Cre recombinase allows for deletion... site of excision, this approach is still not considered safe for therapeutic applications. piggyBac (...insertional mutagenesis, adenoviral vectors are considered a safer way to express reprogramming factors ...many passages. Ban et al developed a temperature sensitive Sendai virus which is removed by culturing cells... regulate Sendai virus transgene expression. Transient episomal delivery Non-integrating methods have...

  11. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    ...capacity of stem cells. Yet, very few researchers consider the sex of their cells when doing experiments—...between male and female cells. The result of not considering the sex of the stem cells was that their male... in Norway, the team realized they should also consider the sex of the stem cells. In this case, they ... many researchers I’ve talked to who’ve never considered the sex of their cells. For one thing, cells ... there are numerous methodological articles to consider—we have loaded those we are aware of in our checklist...clue or potential place to start research is by considering sex disparities in disease severity. Such disparities...checklist. A common worry for researchers wanting to consider sex is cost. Especially in animal research, they...

  12. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Cleavage Assay Not Provided 5% Inexpensive, simple Low-throughput, low sensitivity Sanger Sequencing Provided...inappropriate for heterogenous populations, low sensitivity Targeted Amplicon Sequencing Provided 0.01%...0.01% (variable) Sequence-level data, extremely sensitive Do not sequence all DSBs, may miss unpredicted ...information from targeted amplicon sequencing is highly sensitive with detection levels as low as .01% (Hendel et... with in silico prediction to create a more comprehensive list of off-target editing events. Two of the...laboratories. Nevertheless, unbiased detection isn’t as sensitive as targeted amplicon sequencing. For example, ...throughout the genome. However, their decreased sensitivity means that the best option moving forward may...

  13. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...available with improved kinetic and sensitivity parameters. Sensitivity and Localization The calcium concentrations...are used to monitor calcium concentrations and transients in living cells and organisms. Unlike chemical...kinetics: GCaMPf (fast) GCaMPm (medium), GCaMPs (sensitive but slower). GCaMPf may be suitable for very rapid...You want to use a calcium sensor that is most sensitive to your expected range. For example, calcium in... changes with Ca while another wavelength is insensitive. Additional red sensors based on mRuby (jRCaMP1...unnecessary calcium buffering and to increase sensitivity), and is a great FRET sensor option if you have...integration within the target cells, ensuring more consistent and reproducible results in calcium assays. There...

  14. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...and potential delivery methods.   General considerations for planning your CRISPR experiment As with...experiment, there are many factors that need to be considered during the planning process. For CRISPR experiments... method: Do you need stable integration or is transient expression sufficient? Which cell types will you...are compatible with all your components. Next, consider the best form for those CRISPR components based...selection marker to generate stable cell lines. Transient or stable expression of Cas9 and/or gRNA in a ...transfer vector and used to generate AAV particles. Transient or stable expression of SaCas9 and/or gRNA. Infects...cells using microinjection or electroporation. Transient expression of CRISPR components, expression decreases...

  15. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...showed greater photostability and also matured considerably faster, with expression levels comparable to...PILATeS, Pulling-Induced Luciferase Association Tension Sensor (Zhong et al., 2024). In this new variation...and only exposed when the module unfolds under tension. When this happens, BiT and the LgBiT luciferase... Figure 5: Left: Mechanism of the PILATeS tension sensor. The LgBiT (cyan) fragment of luciferase...protein (purple) only when Titin I10 unfolds under tension. Upon binding, LgBiT and BiT Tag catalyze a luminescent...PILATeS-functionalized coverslip creates mechanical tension, and the luminescence colocalizes with the focal...lower background signal and can be used in light-sensitive systems. Most excitingly, assembly of the split...

  16. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...first-generation APEX suffered from inconsistent activity levels and limited sensitivity under the stringently low...extremely dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP...GFPs with diversity in fluorescence brightness, intensity, excitation and emission spectra. The lab of Dimitri... degradation. Chemical approaches may require expensive peptides or small molecules and exceed a physiologically...physiologically relevant exposure in intensity and/or duration. Both of these strategies carry the risk of...activate signaling cascades on exposure to low-intensity blue light. This approach relies on the incorporation...incorporation of light-oxygen-voltage (LOV) sensing domains from algae into chimeric Opto-RTKs. The LOV domains...

  17. New Norepinephrine Indicators: nLightG and nLightR

    Type
    Blog Post
    ...brain. Sensitivity In ex vivo studies on mouse brain slices, nLightG and GRABNE were more sensitive than...But this first generation of tools had limited sensitivity and speed and were only available as green probes... vivo experiments. nLight indicators are more sensitive to and selective for norepinephrine and faster...that red fluorescent proteins tend to be less sensitive than green fluorescent proteins. nLightR did perform...and may soon have a red tool which matches the sensitivity of nLightG. Many thanks to Mike Waring and Tommaso...Bohacek, J., Reiner, A., … Patriarchi, T. (2023). Sensitive multicolor indicators for monitoring norepinephrine...

  18. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...antibodies when possible. If cost is prohibitive, consider an approach that prioritizes more reproducible...with representative images showing the staining intensity in the KO population (pink histogram) compared...knockout experiments can be time-consuming and expensive, it may be helpful to find an antibody that has...RNA (siRNA). Knockdown caused by siRNA is also transient, and as such, the optimal knockdown of the protein... rate. If you need a more permanent knockdown, consider using shRNA vectors instead. With both siRNA and...understand the relationship between antibody staining intensity by flow cytometry, and mRNA expression.   ... at similar levels in your cell lines, you can consider a cell treatment approach. If there is a cell ...

  19. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ... This is an important downside of the technology and should always be considered when interpreting results...oligomerization of fluorescent proteins is a concern. The propensity of fluorescent proteins to oligomerize is known...brightness, photostability, maturation time, and acid sensitivity of various fluorescent proteins (Botman et al...coded by the multiple cloning site and therefore consist of random amino acid sequences or (ii) designed... an inert, unstructured peptide and therefore consisted of glycines, serines and other small, non-aliphatic...from Aequorea victoria GFP sticks out and can be considered a linker (figure 1). In fact, for several FRET...fluorescent protein fusions There are several choices to consider before generating a fusion protein, e.g. linker...

  20. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... other ligands - almost A to Z! - from ATP to urotensin. The MTRIA sensors will be useful for investigating...displays enhanced brightness, expression, and sensitivity compared with earlier SF-iGluSnFR. Image from... response kinetics and increased brightness, sensitivity, and photostability under two-photon illumination...proteins (miRFP670nano and miRFP720) and the Ca2+-sensing domain from Twitch-2B (Matlashov et al. 2022). ...plasmids here!   Torres Cabán CC, et al. Tuning the Sensitivity of Genetically Encoded Fluorescent Potassium ...LiLac has improved response size and reduced sensitivity to pH and calcium compared to previous lactate...470-495 nm) and is capable of local temperature sensing with sub-millisecond resolution and sub-degree ...
Showing: 161 - 180 of 876 results