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We narrowed to 404 results for: pho;

Showing: 81 - 90 of 404 results

  1. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ... that targets the primary is conjugated to a fluorophore (Figure 1).   Figure 1: Comparison between...exist and you can begin to see why generating fluorophore-conjugated versions for each primary antibody...IgGs and then conjugate them with an array of fluorophores. It is common for labs to keep a few trusted... secondary antibodies conjugated to distinct fluorophores to visualize multiple targets at once. An advantage...to ensure that you are using secondaries with fluorophores whose excitation/emission spectra do not overlap....  Fixation Good fixation maintains sample morphology with minimal impact to the target epitope. There...to the first antibody and is conjugated to a fluorophore. You may need to optimize the concentration and...

  2. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...binding. Photobiotinylation Photobiotin can be used to non-selectively label DNA and RNA. Photobiotin acetate...papers and webpages are listed below: EMSA (Electrophoretic Mobility Shift Assays) ELISA (Enzyme-Linked...consists of a biotin molecule attached to a photoreactive arylazide group. Under strong visible light,...nearby nucleic acids (McInnes et al., 1990). Photobiotinylation is sometimes easier and less expensive than...C., & Symons, R. H. “Preparation and uses of photobiotin.” Methods in Enzymology 184 (1990): 588–600. ...Schwartz, S. A. “Biotinylated probes in the electrophoretic mobility shift assay to examine specific dsDNA...

  3. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...caused partially by fluorophore photoinstability, as well as the tendency of fluorophores to aggregate in ...permit analysis by immunostaining. Although the fluorophores used are fluorescently distinct, their protein...preventing design of antibodies specific to each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained...ideal characteristics (low aggregation, high photostability, and high stability after fixation.) From the...3.0, but with mOrange2, EGFP and mKate2 as the fluorophores; mOrange2 is the default state. In contrast,...cells in which the construct is expressed. The photostability of Brainbow-3.2 is improved due to the addition...

  4. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ...engineering channelrhodopsins to have improved photocurrent properties: The sequence and structures that...channelrhodopsin, as well as predict the channelrhodopsin’s photocurrent strength, wavelength sensitivity, and off-kinetics...predicted channelrhodopsin sequences would have photocurrent properties best suited for less-invasive optogenetics...All thirty were functional and their measured photocurrent amplitude, wavelength sensitivity, and off-kinetics...neurons expressing ChRgers produced stronger photocurrents at low light intensity than existing channelrhodopsins...response. This result showed that ChRger2 stronger photocurrent properties make it better suited for systemic...ChRger2’s improved light sensitivity and higher-photocurrent properties when paired with PHP.eB’s systemic...

  5. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ... fused to the phosphopeptide binding domain FHA. The FHA1 domain binds to phosphorylated PKA substrate...fluorescence intensity due to interactions with photoactivatable fluorescent protein Dronpa. The authors tracked...detect PKA activity can be generalized to measure phosphatase activity, second messenger activity, and protein-protein...peak of 519 nm. These proteins have improved photophysical properties and can be used in quenching, FRET...

  6. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...-fold more photostable and as bright to mVenus, the parent YFP, and is the most photostable YFP to date...improved activity as compared to other recombinase/photodimer pairs. Created with BioRender.com In their... publication, Mary Dunlop’s lab generated a photosensitive Cre, Opto-Cre-Vvd, by splitting and joining...joining the protein regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes....ncbi.nlm.nih.gov/32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin ...contribute to experiments that require high photostability, such as time-lapse imaging or with high light...

  7. Tips for Technical Support Calls

    Type
    Blog Post
    ...I’ve answered hundreds of phone calls and thousands of customer service emails in my six years as a senior...now spend far less time answering help emails and phone calls, I feel compelled to share my years of accumulated...customer, can get the most out of your email or phone call. Though my experience is based solely on my...texting, actually writing an e-mail or making a phone call is a big commitment when you’re busy. Your ...might think. The Addgene scientists answering the phone are smart, but they cannot (and should not) answer... our thread or remove a good 5 minutes from our phone call. If your concern is based on data you have... Abcam and NEB have Ph.D.s answering emails and phones). We can’t be experts on every technique or experiment...

  8. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...cells lines have similar morphologies, being familiar with these morphologies and doing a quick check ...that any medium or washing buffer, such as PBS (Phosphate Buffered Saline), that comes in contact with the...purpose.Learn about the effects of CO2 on sample pH Cell morphology: Shape and size matter You should have a light...incubator). It also helps to get familiar with the morphology of the cell line you’re working with so that ...passaged. This could change their growth rates and morphology, rendering their continued use inadvisable for...passage number. Cells change their growth rate and morphology at high passage numbers (>30-40 times), depending...

  9. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...an opsin (a GPCR) linked to a chromophore and functions as a photoreceptor in the eye. There are two flavors...voltage-gated potassium channels found in Drosophila to phosphatases from sea squirts. This design structurally separates...cell level as they are in vitro. Factors like photobleaching, lack of red-shifted indicators, and brightness...significantly brighter and available with red-shifted fluorophores (find FRET capable GEVIs here!). The only issue...issue with these GEVIs? They perform poorly in two-photon conditions, which are necessary conditions for ...indicator. To give an example, a genetically encoded phosphatase (voltage sensing) hydrolyzes a precursor dye ...

  10. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...all of the fluorophores as it passes through the specimen, which often leads to photobleaching.  Furthermore...focus light. Therefore, there is much less photo-bleaching or photo-damage than in conventional laser scanning...order to improve axial resolution and reduce phototoxicity for imaging live specimens, they combined light...light scattering and absorption.  Finally, the photomultipliers used to detect light in confocal microscopes...obstacles (e.g. air bubbles or high concentration of fluorophores) cast shadows along the illumination path. The...
Showing: 81 - 90 of 404 results