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We narrowed to 404 results for: pho;

Showing: 61 - 70 of 404 results

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

    Type
    Blog Post
    ...Alkaline phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally...accumulation of Pho8Δ60 in the vacuole, where it is cleaved to generate an active enzyme. Pho8 phosphatase activity...protein. To monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in ...Normally PHO8 is synthesized at the ER, delivered to the vacuole via the secretory pathway, and then cleaved...activity or the molecular weight shift of Pho8 from its uncleaved to cleaved form is the final read out of...’s slow and only measures bulk autophagy. Morphological assays Electron microscopy: Electron microscopy...identification of autophagic structures based on morphology. Autophagosomes are relatively easy to identify...

  2. The Fluorescent Vegetables in Aptamer Soup

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    Blog Post
    ... not photostable, and were presented in the study to support the uniqueness of the photostability of the...polymerase III transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11...known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment process used to evaluate... and non toxic to cells. Because it rapidly photobleaches, DFHBI can only be used for qualitative, but...hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO shares many structural... Corn and its small molecule makes Corn more photostable than Spinach and Broccoli, and useful for quantitative...increased brightness compared to Mango, and all are photostable and useful for both live and fixed cell imaging...

  3. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...FSC/SSC), the detected photons can be represented through a graph of photocurrent vs. time. The resulting...tumor-infiltrating lymphocytes of that tumor. In Figure 3 you can see how I gated out my lymphocyte population...(SSC). FSC and SSC depend on the cell’s size, morphology, and structural complexity.   Height, Width...your individual experiment. Practical example: lymphocytes in a mouse tumor sample You made it! So much ...flow analysis, I can however clearly identify my lymphocyte population (10.7% of all recorded events) based...electric potential that can be applied to the photomultiplier inside the cytometer to increase the electric...case, I and other lab members had worked with lymphocytes before, so I knew where to look for them on the...

  4. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ...Tichy et al., 2019), photoswitchable tethered ligands (PTL), and photoswitchable, orthogonal, remotely-tethered...pathways? Here’s where photoswitchable intracellular proteins come into play. Photoswitches can be used to activate... G-protein coupled receptors (OptoXRs), and photoswitches. Light-gated ion channels directly open and ...potential and a host of other cellular functions. Photoswitches are engineered molecules that induce conformational... special, covalently linked molecules (i.e. photoswitches) attached near the binding pocket of an engineered...activates downstream secondary signaling pathways. Photoswitchable tethered ligands hold the ligand in close proximity....   Controlling secondary pathways with photoswitches OptoXRs are great for activating entire signaling...

  5. Antibodies 101: Flow Cytometry

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    Blog Post
    ... levels of the fluorophore or fluorochrome-stained protein will emit a stronger photon than a cell expressing...emits green, yellow and orange photons while PE emits yellow and orange photons. If FITC and PE were being...cannot be used for subcellular analysis such as morphology or subcellular localization studies. For these...light source, a cell expressing the associated fluorophore or stained with the associated fluorochrome, ...captured by detectors. The strength of the emitted photons varies from cell to cell depending on the level...expression, measured using antibody conjugated fluorophores Fluorescent antibodies get around the challenges...contain a mix of cells including granulocytes, lymphocytes, and monocytes. Due to differences in size and...

  6. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...increased brightness, sensitivity, and photostability under two-photon illumination than previous GEVIs. Some...called JEDI-2P (Liu et al. 2022) optimized for two-photon imaging. They use a unique and powerful high-throughput...based on performance metrics specifically for two-photon microscopy. The resulting JEDI-2P has faster response...increased voltage response, brightness, and photostability. It enables sustained deep-tissue imaging in...recording using a fast indicator evolved for two-photon microscopy. Cell. 2022 Aug 16. pii: S0092-8674(...domain of ZAP70 (ZtSH2) which binds only the phosphorylated state of an immunoreceptor tyrosinase-based...only when the RTK is activated and the ITAM phosphorylated. To demonstrate that pYtags can be generalized...

  7. 15 Hot Plasmids from 2017

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    Blog Post
    ...to our mScarlet Podcast Segment    Photocleavable protein PhoCl expands optogenetics toolbox Increasingly...proteins. PhoCl is a new member of this class that works via a different mechanism - the PhoCl protein ...illustrates the utility of PhoCl in a variety of settings. For instance, inserting PhoCl in between a protein... al., 2007) and includes four promoters, six fluorophores with nonoverlapping emission spectra (N- and...initiation of reprogramming (evaluated by an alkaline phosphatase-positive assay) as well as stabilization (Nanog-GFP...mScarlet-H with M164H (2-fold improvement in photostability compared to mScarlet, fluorescent lifetime ...NES, enables control over localization. Inserting PhoCl between an enzyme and an inhibitor domain similarly...

  8. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ...salts. Capillary electrophoresis Quantifying DNA using capillary electrophoresis is similar to quantifying... using gel electrophoresis. Except, it’s smaller. And automated. Unlike gel electrophoresis, you only ...wavelength that they maximally absorb. The spectrophotometer measures these absorbances using UV-transparent...specific for double strand DNA (as compared to spectrophotometric methods that measure all nucleic acids). ... sample concentration for you. Agarose gel electrophoresis Though not the quickest way to quantify DNA...methods can’t tell you and you don’t need a spectrophotometer or a fluorometer to quantify DNA this way....nanofluidic channel and the sample is separated by electrophoresis. Smaller fragments migrate quicker than larger...

  9. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    Blog Post
    ...can hydrolyze phosphatidylcholine (PC), the predominant membrane lipid, to phosphatidic acid (PA) or it...activity. Researchers have designed a variety of photoswitchable elements into many other types of proteins.... of experiments. Opto-RhoGEFs manipulate cell morphology and adhesion But what if you’re interested in...cytoskeleton – actin? Actin dynamics control cell morphology, movement, and more, often driven by Rho family...filopodia formation, cell contraction, or other morphological changes, depending on the specific Rho GTPase...published OptoPLD, an optogenetically controlled phospholipase D (PLD). Now they present a set of variants ... it can swap phosphatidylchonine’s choline head group for other natural or synthetic head groups. The ...

  10. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ...available now. It is for recombinant expression of phosphoserine-including proteins in E. coli (Zhu et al., 2022... 2022). It uses the TAG codon for the phosphoserine and, because it uses ∆serB strains, does not require...require exogenously added phosphoserine. A community resource Like Addgene, GCE4All is a community resource...shortlisted several protocols, including their phosphoserine ncAA protocol, to optimize and share, they’re..., R. (2022). Site-specific Incorporation of Phosphoserine into Recombinant Proteins in Escherichia coli...
Showing: 61 - 70 of 404 results