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We narrowed to 400 results for: pho;

Showing: 91 - 120 of 400 results

  1. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ... Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is the first step of a western. ...s post-translational modifications such as phosphorylation, acetylation, methylation and ubiquitination...ubiquitination. For example, to test if a protein is phosphorylated following a specific experimental condition you....2015.04.012 Burnette WN (1981) “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide....76.7.3116 Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels...

  2. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...identification of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique that can detect... detect multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene...Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med. 2020;20(10):798-805. doi: 10.2174... He Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence of plasmid ...plasmid DNA in untreated capillary. Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/elps.201000061. PMID...topoisomers, knots, and catenanes by agarose gel electrophoresis. Methods Mol Biol. 2009;582:11-25. doi: 10.1007...topology analysis of plasmid DNA by capillary gel electrophoresis. BioProcess Int. 2006 Sep;4(8):38-40. Summers...

  3. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ... construct for every application. Plus, the photophysical properties of chemical dyes provide many advantages...strategies. The green structure represents a fluorophore or other useful ligand, gray structure is the...non-natural amino acids to directly incorporate a fluorophore in your protein or to incorporate an alkene to...Organic dyes are typically both brighter and more photostable than fluorescent proteins (often up to 10-fold...years; improving the labeling kinetics or the fluorophore’s microenvironment to enhance brightness, stability...Better Dyeing Through Chemistry & Small Molecule Fluorophores. Addgene Blog. https://blog.addgene.org/better-dyeing-through-chemistry-small-molecule-fluorophores...better-dyeing-through-chemistry-small-molecule-fluorophores Los, G. V., Encell, L. P., McDougall, M. G.,...

  4. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...signals upon sensing glutamate using both 1-photon and 2-photon imaging.   Figure 1: 1P and 2P excitation...membrane trafficking and localization. Improved photophysical properties The improved variants of iGluSnFR3...pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol anchor. SGZ contains a PDGFR transmembrane...

  5. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    ...Xin Zhou greatly expanded that list by using photoswitchable fluorescent proteins to create enzymes that...turned the enzymes back on. Xin termed these photoswitchable enzymes Fluorescent Light-inducible Proteins...various designs, Xin has been able to develop photoswitchable Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5,... You can learn more about Xin’s work with photoswitchable components of the ERK signaling pathway in ... the production of HMOs. Current HPLC-MS and fluorophore-based technologies are too time consuming in ...Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins Use Light to Control ...

  6. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    ...envelope, Phoenix-ECO, (for infection of mouse and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for...of the viral packaging cell line. For example, Phoenix, a second generation γ-retrovirus packaging cell... produced in just a few days. Another variant, Phoenix-gp, contains only Gag-Pol and enables additional...

  7. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ... efficiency by which absorbed photons are converted into emitted photons. These are indicated by the extinction...this series of posts are (i) brightness, (ii) photostability and (iii) aggregation tendency. Brightness...high brightness is an important feature of a fluorophore. The key question is how the brightness is best...Theoretical brightness Two key properties of fluorophores that determine brightness are the extent to ...Without further information about properties like photostability or aggregation tendency, our choice would be...

  8. Participate in the Pets of Addgene Scavenger Hunt

    Type
    Blog Post
    ...clue 1 below!) You will find five photos this way. At the very last photo, you will find a link to a form...workspace over the last year. Now, we’ve collected photos of our pets with Addgene gear, and have unleashed...Addgene-branded clothing on the blog. Below each photo, the pet gives you a clue to help you find the next... next picture. If you find the next correct photo, you will see a reference to this scavenger hunt and...

  9. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...et al., 2013), incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red) to solve...expect to see systems that use newer, more photostable fluorophores and that can be combined with new imaging...their technology Brainbow. Brainbow 1.0 used the fluorophores RFP (red), YFP (yellow), and membrane-tethered...clearing methods and can be labeled by synthetic fluorophores, which are more stable than fluorescent proteins...

  10. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...of which you can find at Addgene. Biliverdin chromophore Near-infrared fluorescent proteins (NIR FPs) ...) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically. Fortunately...from your source and to your camera.  Two-photon image of miRFP670nano3-expressing neurons in the... derived from bacterial phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than...were derived from cyanobacteriochrome (CBCR) photoreceptors (Oliinyk et al., 2019). They were engineered...Molecular brightness vs. iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells...

  11. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...inactivate the fluorophores in the sample, either by chemical treatment or photobleaching or by stripping...and that each secondary has a different color fluorophore that you can image separately on your microscope...labeling sequentially with one, inactivating the fluorophores, then labeling with the other, you can still...using primary antibodies that are conjugated to fluorophores. However, the primary-secondary workflow (a....fluorescently-conjugated streptavidin to get more fluorophores per antibody, or conjugate with an enzymatic...

  12. In Living Color: The Skinny on In Vivo Imaging Tools

    Type
    Blog Post
    ...last year, Verkhusha added two far-red light photoactivatable (PA) near-infrared fluorescent proteins (FPs...fluorescent proteins, iRFP670 and iRFP720, as photoacoustic contrast agents for two-color imaging in animals...genetically-encoded probes by making them brighter, more photostable, smaller, and less toxic to cells. They’d like...751-4. doi:10.1038/nmeth.2521) Far-red light photoactivatable near-infrared fluorescent proteins engineered...: 10.1038/ncomms3153. PubMed) Multicontrast photoacoustic in vivo imaging using near-infrared fluorescent...

  13. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...for up to 60% of the cell culture contamination (Uphoff, 2002). Mycoplasmas are considered to be one of...from lowered pH due to microbial contamination. Photo by Kaustubh Kishor Jadhav. Mycoplasmas can remain...for at least two or three days before sampling (Uphoff and Drexler, 2011). Culturing on agar plates/broth...works in the tissue culture hood while wearing PPE. Photo by Kate Harten DeMaio. Cell lines and reagents... of mycoplasma species. J Microbiol .44: 42–49  Uphoff Cc, Drexler Hg (2002) Comparative Antibiotic Eradication...10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma Contamination...

  14. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore (circularly...three-dimensional cultures that mimic aspects of organ morphology and physiology in vitro and have emerged as a...investigating genetic mechanisms related to morphogenesis, cell fate specification, and human disease....relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity, ...

  15. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...signal-to-noise ratio Less chance of phototoxicity Reduced photobleaching Simplified, long-term single-molecule...enough signal. This workaround risks cellular phototoxicity and eliminates the possibility of long-term ...a higher signal-to-noise ratio with a lower photobleaching rate. Given the power of SunTag, they attempted...Protein Oligomerization Learn about Small Molecule Fluorphores Resources on Addgene.org Find Fluorescent Protein...

  16. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...contributed by Leah Schwiesow Calcineurin is a protein phosphatase that responds to rising intracellular calcium... of Ca2+ are high in the cell, calcineurin dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent... intercalating fluorescent dyes which can cause photo-induced DNA breakage and structural deformation,...reversible and do not cause structural deformation or photo-induced damage. These fusions can also function ...

  17. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...fluorescent protein joins mTFP1 and mWasabi, as fluorophores from Allele Biotechnology that now can be deposited...brightest teal fluorescent proteins that were photostable and had a high 480/530 nm emission ratio. They...mWasabi is 1.6-fold brighter than EGFP and is photostable. It also has narrow excitation and emission peaks...useful in multicolor labeling particularly with fluorophores that excite in the violet range. For example...Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein...

  18. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...migration of a cancer cell line equipped with photodissociable EB1 could be stopped in areas with blue light...manipulating and studying cellular dynamics. The photoinducible control of protein function is more specific...signal. Prior to the development of these probes, fluorophore binding aptamers and MS2-binding fluorescent ...contain a smaller RNA tag, are less prone to photobleaching, and achieve better signal in vivo as compared...aptamers or MS2 tags using the same number of fluorophores. You can find plasmids containing the cobalamin...

  19. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...sfCherry2 sfCherry2(1-10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1-10) Improved split...iRFP713. Requires presence of heme oxygenase for chromophore formation (abundant in eukaryotes) A Near-Infrared...domain. Requires presence of heme oxygenase for chromophore formation (abundant in eukaryotes) Bright monomeric...typically irreversible and requires some time for chromophore maturation, splitFAST displays rapid and reversible...flexibility in color labeling with exogenous fluorophores, which can even be bright enough for single-...

  20. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ... molecules, which are much brighter and more photostable than fluorescent proteins, making them ideal ...versions. ASAP5 can be imaged using one- or two-photon microscopy to detect action potentials and subthreshold...fluctuations in awake behaving mice through two-photon microscopy at depths up to 600 µm. Figure...recognizes the disease-associated version of Tau (phospho-Tau). We hope these will become valuable tools ...

  21. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...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...

  22. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can... the instability of the supposedly alien 5’-phosphorylated ssDNA guides, which might be rapidly degraded...things (101 lessons on Spanish, C++, guitar, photography) and often dreams of a sunny day in Edinburgh...

  23. DNA Purification Without a Kit

    Type
    Blog Post
    ...molecules and coaxes it into sticking to the hydrophobic glass filter. Each glass filter can capture up... only requires incubating the columns with 1M phosphoric acid for 3 minutes followed by a short centrifugation...purification columns could also be cleaned with 1M phosphoric acid and re-used, but trace contamination could...

  24. DNA Service - Cloning Grade DNA

    Type
    Collection
    ... and 260/280 ratio has been determined by spectrophotometer. Other methods of quantification may yield...

  25. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ... Figure 1: Plasmid mix to label neuronal morphology (eYFP) and the synaptic protein PSD95 (PSD95-...control   Figure 2: Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry...experiment it is possible to compare neuronal morphology and synaptic connectivity before and after the...Micron Plasmid Attaches Covalently to DNA via a Phosphotyrosyl Linkage.” Molecular and Cellular Biology 5(...

  27. A Control for All Seasons

    Type
    Blog Post
    ...well-studied kinase known to prevent tau hyperphosphorylation. You could run brain samples from both groups... molecular weight shifts indicative of tau phosphorylation. By including the positive control, you will...controls are single stained samples for each fluorophore and are critical to analyze multiplex flow cytometry...

  28. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ... the phosphatase you choose. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase) are ...overhangs after digestion, you will need to use a phosphatase to prevent re-circularization of the backbone... treat your digested backbone plasmid with a phosphatase prior to the ligation step or prior to the gel...

  29. Editor's Choice, July 2016

    Type
    Blog Post
    ...CRISPR variant going forward.   Special Delivery Fluorophore Targeting for FRET Studies While I love all of...This post describes how to target and use FRET flurophores for a variety of applications. While editing ... 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...

  30. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...et al. "A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas...Deisseroth, combined optical and genetic methods to photo-stimulate mammalian neurons and alter synaptic events...rhodopsins, light-sensitive proteins that absorb photons for energy conversion or the induction of intra...resulting viral-infected neurons are subsequently photo-stimulated through fiberoptic cannulas directly ...
Showing: 91 - 120 of 400 results