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... 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...

...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...

...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...

... 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...

...-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...

...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...

...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 ...

...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...

...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...

...confocal and widefield imaging, two-photon microscopy, and fiber photometry. Plus, both designs offer promising...had better brightness, signal-to-noise ratio, photostability, and signal response upon Ca2+ binding. Most...importantly, it doesn’t display the problematic photoswitching upon blue illumination that plagues other red...mApple, which means it likely has the blue-light photoswitching issue). Although similar in brightness to jRGECO1a...calcium sensor with exceptional brightness, photostability, and multiplexing capabilities. bioRxiv 2024.12.16.628673...incubate at room temperature, and image with smartphones. The Qi lab also developed an algorithm called...

... 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...

...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...

...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 ...

...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...

...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...

... 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...

... 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....

...Here's what you'll find in this post: Bright and photostable yellow fluorescent protein mGold2 Viral delivery...synaptic connections with ATLAS Bright and photostable yellow fluorescent protein mGold2 By Mike Lacy... current yellow fluorescent proteins tend to photobleach rapidly, which hinders their use for long-term... The mGold2 variants significantly enhanced photostability — 25-fold over mVenus and 4-fold over the previous...mGold2s, or mGold2t. Bottom: Quantification of photostability. Image reused from Lee & Lai et al. (2025) ... Lee, J., Lai, S., et al. (2025). Bright and photostable yellow fluorescent proteins for extended imaging...

...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...

...more! Read on to learn more about: Highly photostable mScarlet3 variants Addgene's own AAV2 and AAV5...brightest red fluorescent protein, but its modest photostability limits applications like long-term imaging ...mutagenesis and screening strategy to select photostable variants, resulting in mScarlet3-S2 (M163H/M66Q...mScarlet3-H showed up to 29- and 15-fold higher photostability than mScarlet3, respectively. Although they...comparable or a bit better than mCherry), their photostability means they more than make up for it over the...Wang, K., Xue, F., … Xu, P. (2026). A highly photostable monomeric red fluorescent protein for dual-color...