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We narrowed to 404 results for: pho;

Showing: 71 - 80 of 404 results

  1. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...categories of fluorescent proteins Photoactivatable / Photoconvertible: These proteins can switch their ... fluorescence. Examples for such photoactivatable or photoconvertible proteins are PA-GFP, Dendra2, and...e.g. mCherry). Oxygen: The maturation of the chromophore on many FPs (particularly those derived from ...takes the FP to correctly fold and create the chromophore. This can be from a few minutes after it is translated...0.1) and this should be taken into account. Photostability can be affected by experimental parameters ...excitation light intensity, pH or temperature). Photostability: Fluorescent molecules gets bleached (i.e. ...after prolonged exposure to excitation light. Photostability can be as short as 100ms (EBFP) or as long ...

  2. Hot Plasmids - October 2022

    Type
    Blog Post
    ...absorption of 540 nm and two-photon excitation around 1,040 nm, has larger photocurrents, and is more selective...they found an indicator that is optimized for 2-photon imaging of voltages. This includes monitoring of...resultant JEDI-2P sensors overcome many challenges in 2-photon in-vivo imaging of voltage fluctuations. Image ...recording using a fast indicator evolved for two-photon microscopy. Cell, 18, 3408 - 3425.e29. (2022). ...channelrhodopsins (KCRs), from a fungus-like protist Hyphochytrium catenoides (Govorunova et al. 2022). The authors...have high selectivity for K+ over Na+ and rapid photocurrent kinetics. They demonstrate the use of HcKCR1...frequencies with (magenta) and without (black) photoactivation of HcKCR1 in mouse neurons. D) Membrane voltage...

  3. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ...panel.  First, fluorophores vary in brightness. You don’t want a super-bright fluorophore, like PE, on ...assay you can do. Most non-HRP conjugates are fluorophores, which work not by reacting to a chemical, but...then read by a detector panel in the machine. Fluorophores are light sensitive, so antibodies conjugated...before you make your purchase. Picking a single fluorophore for an experiment is easy, but what about those...of interest! Research the brightness of your fluorophores before buying and try to inversely pair brightness...possible.      Fig 2: Try to inversely pair fluorophore strength with protein abundance.    Second...Second, you’ll need to consider spectral overlap. Fluorophores signal are actually quite broad, resulting in...

  4. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ... As Nano-lanterns are not subject to photobleaching/phototoxicity, they successfully imaged multiple subcellular...long-term imaging, especially where photobleaching and phototoxicity are a concern, these constructs come...Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be affected by...by both autofluorescence and photobleaching. Researchers have long been interested in using luminescence...bioluminescence resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant are used...in the culture medium to allow RLuc8 to produce photons. The original nano-lantern emitted yellow-green...Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check Out Fluorescent...

  5. Antibodies 101: Conjugation

    Type
    Blog Post
    ...together. Common conjugates in the lab Fluorophores Fluorophores allow you to detect your antibodies using...microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available for antibody conjugation...tool for comparing different fluorophores. We also touched on fluorophores as antibody conjugates in an...sure to check the spectral properties of the fluorophores you are considering to make sure that they’ll...equally, if not more, common conjugate class than fluorophores are enzymes. The most common enzyme conjugates...are Horseradish Peroxidase (HRP) and Alkaline Phosphatase (AP). Various substrates for each of these enzymes...

  6. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...enzymes that seal breaks in DNA by joining 5 ́-phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In...ligation product in a gel, but we can now use fluorophore- quencher pairs or qPCR-based methods to greatly...downstream side (the base pair providing the 5 ́-phosphate to the ligation). The structural/mechanistic reason...the reaction. This “peeling back” of the 5 ́-phosphorylated base can be observed in the crystal structures...identifiable and quantifiable by capillary electrophoresis (CE) (see below). This method allowed screening...gene PCR-ligase detection reaction-capillary electrophoresis assay." Journal of clinical microbiology 45.6...mixed population of excess wild‐type DNA." Electrophoresis 34.9-10 (2013): 1415-1422. PubMed PMID: 23463388...

  7. Science as Art

    Type
    Blog Post
    ...scientist and photographer based at the University of Queensland, whose outstanding marine photography has attracted...” Daniel went on to discuss his work: “Macro photography has been my passion for a while - a tool to expand...life cycle of the creature they deal with.” So photography can be used as a way to better understand and...scientist and non-scientist alike. “The other thing photography has shown to me - it requires an awful amount...data, publish in time, defend the thesis, etc. Photography is critically important in many fields of biology... can be found on his website http://www.microworldsphotography.com/. Another artist who spent his life...artists can compete in the Nikon Small World Photomicrography Competition - a 40 year old international ...

  8. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...mEGFP (Zacharias et al., 2002). GFP needs oxygen Chromophore formation requires a fully folded beta-barrel...an intramolecular reaction that generates the chromophore (Tsien, 1998). Importantly, this reaction requires...-by-case basis. GFP is sensitive to acid The chromophore of GFP can exist in a protonated and a deprotonated... bind specifically and with high affinity to fluorophores. Examples are the FlAsH tag, the His tag and...enzyme to covalently label the peptide with a fluorophore. Both systems are a hybrid between a genetically...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains...

  9. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...yourself or you could “clone by phone”. One of my professors was excellent at phone cloning with labs around ... NanoLuc® Luciferase from a rather dull 19kDa Oplophorus luciferase by Promega’s Advanced Technologies...problematic because excitation light can cause phototoxicity, autofluorescence, and perturb biological phenomena... molecules. FRET sensors face challenges of photobleaching, autofluorescence, and, in the case of exciting...exciting cyan-excitable donors, phototoxicity. Another challenge to using FRET sensors comes when employing...sensor with the following characteristics: No photoexcitation High Ion specificity High sensitivity Easy ...and colleagues (8) were interested in using photoactivatable, optogenetic accuators to depolarize or hyperpolarize...

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

    Type
    Blog Post
    ...spectrum (hence the name), which is due to a chromophore formed from a maturation reaction of three specific...surprising aspects of GFP was the fact that the chromophore forms spontaneously and without additional co-factors... β-sheet-containing “barrel” shape, with the chromophore concealed at the center of the structure, shielded...that increased the fluorescence intensity and photostability of GFP. This also shifted its major excitation...been introduced to GFP and new iterations of fluorophores are constantly being engineered. Table 1 lists... 1: The specific mutations comprising common fluorophores Fluorescent Protein Mutations Relative to...Known Function S65T Increased fluorescence, photostability, and a shift of the major excitation peak to...
Showing: 71 - 80 of 404 results