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We narrowed to 78 results for: fluorophore

Showing: 1 - 20 of 78 results

  1. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ... you'll need to pick a fluorophore for each target and ensure the fluorophores, together, are able to ...outlining a few commonly used fluorophores and their emission spectra. Some fluorophores carry their emission ...and RB545. Fluorophores can also come in combination of two conjugated single fluorophores, like APC-Cy7...respective colors of a few commonly used fluorophores. Each fluorophore has a defined emission maximum. Note...individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores. A fluorophore's...voltage, the concentration of the fluorophore, and whether that fluorophore appears alone or in conjugation...conjugation with another fluorophore. The brighter a fluorophore, the better the signal-to-noise ratio, and the...

  2. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...each sample has a limited number of fluorophores and each fluorophore can emit a limited number of photons...beholden to chemists and (expensive) synthetic fluorophores. Add a dash of DNA with an electrical jolt and...cells become perfectly capable of synthesizing fluorophore fusions on their own. Subsequent advances in ...gene-edited cells can decrease the number of fluorophores, compromising the photon budget. Likewise, the...extract a few more photons from a sample. Chemical fluorophores can be substantially brighter and more photostable...with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that binds to the tag. Like...a small substrate ligand motif attached to a fluorophore (Figure 1b). Engineered ligases (e.g., lipoic...

  3. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ...to identify a fluorophore, spectral flow cytometers use all detectors for all fluorophores. This allows...samples stained with a single fluorophore. They’re required for every fluorophore you use in your panel, whether...conventional flow cytometry, light emitted by a fluorophore is guided to a single detector using a series...flow cytometry uses one detector to detect one fluorophore and therefore one cellular marker. However, emission... emission spectra of certain fluorophores can “spill over” into detectors that are not the primary detector...detector for that fluorophore. For example, in figure 1, the FITC signal is spilling over into the primary...the full spectrum of light to distinguish one fluorophore from another. Instead of using a single (primary...

  4. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...donor fluorophore to the acceptor fluorophore. The typical steps in FRET are: 1. Donor fluorophore excitation...acceptor fluorophore--think of it as a virtual photon 3. Relaxation of the acceptor fluorophore back to...acceptor fluorophores must be compatible, such that the emission spectrum of the donor fluorophore overlaps...typical excitation and emission of an excited fluorophore, FRET involves a non-radiative transfer of energy...with the excitation spectrum of the acceptor fluorophore. Otherwise the energy transferred from the donor...addition to sufficient spectral overlap, the fluorophores must be located within 1-10nm of each other ... is used to infer an interaction between the fluorophores and serves as a small-scale ruler to measure...

  5. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...non-radiatively from an excited molecular fluorophore (the donor) to another fluorophore (the acceptor) by means of ...the two fluorophores used are in close vicinity. Thus, the distance between the two fluorophores and their... of a fluorophore pair, donor:acceptor stoichiometry, and cross-talk between the two fluorophore colours...dipole-dipole coupling." The emission of the acceptor fluorophore can be measured using microscopy techniques. ...interactions within living cells. By coupling fluorophores to proteins, pioneers of this technique were...a biosensor, as the distance between the two fluorophores and their spatial orientation will be fixed.... a given wavelength, to the concentration of fluorophore in solution) are two additional parameters that...

  6. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...biophysical interaction between two fluorophores. When the first fluorophore (the donor) is excited, it can...acids/lipids of interest are fluorophore labeled and that these fluorophores can undergo FRET when they ...targeting a FRET fluorophore to a biomolecule is a bit like a Fedex delivery. Your fluorophore package needs... energy directly via resonance to the second fluorophore (the acceptor) if the two probes are close together...physical distance between the donor/acceptor fluorophores (Stryer, 1978), a measurement that is unaffected...label your biomolecule of interest with FRET fluorophores? When is absolute labeling specificity needed...measure it). Molecular mailbox: How to get your fluorophore delivered to the right place? FRET experiments...

  7. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ...state contains a fluorophore and a quencher in close proximity. For instance, the fluorophore and quencher...of aptamers means they can be used with or as fluorophores in several different ways. Researchers have ...fluorescent in situ hybridization (FISH) where fluorophore-linked antibodies are used to visualize RNA. ...removing the quencher from the proximity of the fluorophore and subsequently leading to a positive signal...signal. The same effect can be achieved for a fluorophore that relies on dimerization for activation, only...used as powerful biosensors, the bound protein fluorophores and quenchers are still relatively bulky. One...fluor. Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether...

  8. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ... conjugated to a fluorophore, not every marker will have a corresponding fluorophore-conjugated antibody...cases, fluorophore-conjugated antibodies may exist, but only with a limited selection of fluorophores. If... tag but not a fluorophore. In a separate step, cells are incubated with a fluorophore-conjugated streptavidin...wide range of fluorophores, allowing for great flexibility when it comes to fluorophore selection for ...common method is a cell surface label, where fluorophore-conjugated antibodies directly bind to epitopes...employ beyond simply incubating cells with single-fluorophore-conjugated antibodies. These methods can include...interaction (Figure 1a). This interaction brings the fluorophore in proximity to the target marker, allowing for...

  9. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ...function in live cells. Fluorophore properties: The specific properties of the fluorophore, such as excitation...ligands Figure 1: Example structures of fluorophores, fluorescent dyes, and fluorescent ligands. ...refers to the emission of light by a substance (fluorophore) that has absorbed light or electromagnetic radiation...another wavelength (Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ...emission spectrum. Fluorescent dyes utilize these fluorophores by combining them with a molecular species with...such as proteins, nucleic acids, or glycans. Fluorophores can also be covalently attached to self-labeling...a small substrate ligand motif attached to a fluorophore. When paired with a protein of interest, these...

  10. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    Published
    Feb. 1, 2022, 2:15 p.m.
    ...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...

  11. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...together. Common conjugates in the lab Fluorophores Fluorophores allow you to detect your antibodies using...tool for comparing different fluorophores. We also touched on fluorophores as antibody conjugates in an...cytometry, or spectrophotometry. The range of fluorophores available for antibody conjugation spans the...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...

  12. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...Photobleaching is the irreversible destruction of a fluorophore under the influence of light. Any fluorescent... photostability The photobleaching rate of a fluorophore primarily depends on the excitation power and...and the excitation wavelength. A fluorophore is less likely to bleach when it is excited off the peak wavelength...unit of time, decreasing the likelihood that a fluorophore will bleach. This is, however, not the full story... a brief time), whereas in widefield imaging fluorophores are excited with relatively low light levels...intensity over time. In order to predict how fluorophores perform in ‘real experiments’, it is advisable...cases, the fluorescence intensity decay of a fluorophore does not follow a simple mono-exponential decay...

  13. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ... 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...to the first antibody and is conjugated to a fluorophore. You may need to optimize the concentration and...your primary antibody and 2) is conjugated to a fluorophore that suits your experiment (i.e. the spectral...

  14. Editor's Choice, July 2016

    Type
    Blog Post
    Published
    Aug. 5, 2016, 3 p.m.
    ...CRISPR variant going forward.   Special Delivery Fluorophore Targeting for FRET Studies While I love all of... to study biological phenomena and about how fluorophores other than fluorescent proteins can be incredibly...Comparison to Cas9 and NgAgo Special Delivery: Fluorophore Targeting for FRET Studies Gendered Innovations...

  15. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...started!   Thanks for all your hard work!   Fluorophores When is a Monomer not a Monomer? The Top Three...Better Dyeing Through Chemistry & Small Molecule Fluorophores In this post, Luke Lavis from Janelia Research...fluorescent proteins are incredibly useful, chemical fluorophores can still be used in a wide variety of experiments...experiments. Read more to find chemical fluorophores for your next experiment. Nucleic acid based machines...

  16. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment process used to evaluate...hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO shares many structural...In addition to these established RNA aptamer fluorophores, more are in development, including Carrot and...Dolgosheina, Elena V., et al. "RNA mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling...polymerase III transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017...on the Addgene blog Learn more about aptamer fluorophores Investigate other fluorescent biosensors Find...

  17. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ...example, if your flow cytometer reads out the fluorophores FITC and PE on the same laser, you will need...deciding to use them, using a tool like BioLegend’s Fluorophore Guide Chart.  Pro tip! Some antibodies go by ...change the compensation values for whichever fluorophore is present in the sample (APC in the case of ...other channels you’re using first. A different fluorophore channel may be affecting your populations and... this population is double-positive for both fluorophores and that there is no issue with compensation...

  18. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    Published
    Oct. 10, 2017, 1:57 p.m.
    ...background. In addition to this, the use of multiple fluorophores can add a second layer of contrast, color contrast...further exploit the unique characteristics of fluorophores to block out of focus fluorescence from reaching...large number of optical slices are acquired all fluorophores throughout the sample are constantly exposed...sample via an evanescent wave that only excites fluorophores within a few hundred nanometers of the glass...TIRF) microscope can be used. TIRF only excites fluorophores within a few hundred nanometers of the coverslip...contrast by filtering out emission light from fluorophores higher in the sample. TIRF is also very gentle...the sample and excitation occurs through the fluorophore’s interaction with an evanescent energy field....

  19. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore (circularly...relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity, ...

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

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...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...3.0, but with mOrange2, EGFP and mKate2 as the fluorophores; mOrange2 is the default state. In contrast,...
Showing: 1 - 20 of 78 results