...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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, ...

...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...

...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 ...

...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...

...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...

...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-...

... 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 ...

...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...

...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...

...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...

... 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(...

... covered in this segment Hot Plasmid 1: Photocleaveable PhoCl protein for controlling molecular processes...processes with light. Listen to Our Segement on PhoCl Segment  Hot Plasmid 2: Ouabain selection for increased... phocl...