We narrowed to 507 results for: otos
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TypeBlog PostPublishedAug. 9, 2018, 1:04 p.m....well-established protocols for their production and purification that yield high titers (see also AAV protocols for...neuronal connectivity. These methods are less neurotoxic and better compatible with other neuroscience...system where it replicates, spreads, causes neurotoxicity, and is lethal if left untreated (for review...AAV), have been used for tracing and visualizing motor, visual, and auditory neuronal circuitry in vivo... project but neurons were traced with AAV2. Two-Photon imaging was used to generate a 3D finemap of neurons...WGA-Alexa Transsynaptic Labeling in the Phrenic Motor System of Adult Rats: Intrapleural Injection versus....M., and Mooney, R. (2017). Identification of a motor to auditory pathway important for vocal learning...
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Hot Plasmids: Summer 2025
TypeBlog PostPublishedAug. 28, 2025, 1:15 p.m....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...production processes. Check out our step-by-step protocols for AAV production, purification, and titration... -
New Educational Resource: The Addgene Videos Page
TypeBlog PostPublishedJune 16, 2017, 2 p.m....sections: Protocols To complement our written protocol materials, we have also created video protocols. These...navigate various lab processes. Our most recent protocol video detailing Streaking Plates for Single Bacterial...Check Our Our Protocol Pages Browse our Plasmid Reference Pages ... protocol video... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog PostPublishedNov. 21, 2024, 10:47 p.m....optogenetics — and shared the protocols we use in our own labs. By 2023, we had 47 protocols and 12 guides available...experiment. Table 1: Most popular guides, protocols, and collections of 2023. Educational Resource...Guide Optogenetics Guide γ-Retrovirus Guide Protocols Agarose Gel Electrophoresis Bacterial Transformation...around the world. Collectively, our 12 guides, 47 protocols, and 186 collection pages received over two million...a thousand words, especially when it comes to protocols. In 2013, we launched the Addgene YouTube channel...Figure 4). Four of the five most popular videos are protocol videos, where showing exactly how to execute a... -
CRISPR/Cas9 FAQs Answered!
TypeBlog PostPublishedMarch 13, 2014, 4:08 p.m.... compared to where the target site sequence (protospacer+PAM) is located? If there are other PAMs in the... there needs to be extensive homology of the protospacer followed by PAM. See Addgene's CRISPR resources.... In the case where you have intact CRISPR protospacer target in the HR template, you usually would ...make sure it's a silent mutation). Q8: Is a protospacer followed by the PAM (my target sequence) in my...template an issue? A8: Yes. If you have intact protospacer+PAM sequence within the HR template, it will ...around 50 µl of the extraction solution, runs the protocol, then uses around 2–4 µl in each 50 µl PCR reaction...engineering using the CRISPR-Cas9 system. Nature Protocols, 8(11), 2281–2308. https://doi.org/10.1038/nprot... -
Exploring Applications of the Bioluminescent HiBiT Tag
TypeBlog PostPublishedOct. 2, 2025, 1:15 p.m....trafficking and their detection can suffer from photobleaching and cellular autofluorescence (Tsien, 1998)...plates or media, which can obscure weak signals; photobleaching from the excitation light, causing the signal...generating reactive oxygen species that can lead to phototoxicity, perturb biological processes, and compromise... Kamio, T., Kono, Y., Hirosuna, K., Ozato, T., Yamamoto, H., Hirasawa, A., Ennishi, D., Tomida, S., Toyooka...Reynaud, E. G., & Shroff, H. (2017). Assessing phototoxicity in live fluorescence imaging. Nature Methods... -
Antibodies 101: Multiplex Immunofluorescence
TypeBlog PostPublishedMay 23, 2023, 1:15 p.m....Frozen Tissue-V2 v2. protocols.io. https://doi.org/10.17504/protocols.io.bs68nhhw Im, K., Mareninov, S...antibodies available at Addgene Protocols for Immunocytochemistry Protocols for other antibody applications...the sample, either by chemical treatment or photobleaching or by stripping out the antibodies, then start...the nuclei. (See Brewer et al, 2022 for a sample protocol.) Simultaneous IF Depending on your needs, your... separately on your microscope. Abcam's sample protocol is a good place to get started. One important... -
Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility
TypeBlog PostPublishedMay 18, 2018, 7:32 p.m....foundation to begin experimenting. Protocols.io, JoVe, and Bio-protocol all work to provide more descriptive...Researchers should be able to follow one another’s protocols and use one another’s reagents with the confidence...that aren’t always mentioned in static written protocols. It is therefore a bit ridiculous to expect that...to share in-depth information about reagents, protocols, analysis tools, and more. Being that this is ...descriptive and interactive protocols that make it much easier to learn new experimental techniques. Electronic...reproducibility List of Tools for Reproducibility at Protocols.io Additional Resources on the Addgene Blog FPbase... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog PostPublishedFeb. 8, 2018, 12:17 p.m....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... -
Antibodies 101: Fc Effector Functions
TypeBlog PostPublishedMay 18, 2023, 1:15 p.m....Antibody dependent cellular cytotoxicity (ADCC) Antibody dependent cellular cytotoxicity (ADCC) is a slightly...deposition (ADCD) b) antibody-dependent cellular cytotoxicity (ADCC) c) antibody-dependent cellular phagocytosis...expressing Fcγ receptors binds to IgG and release cytotoxins that kill the target cell. If the pathogen is...and eosinophils, where the eosinophils release cytotoxins via degranulation. Antibody-dependent cellular...JV. Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor targets. Nat Med. 2000 Apr;6(4... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog PostPublishedOct. 17, 2019, 12:28 p.m....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 ...modification technology in the mouse." Current protocols in immunology 105.1 (2014): 10-34. PubMed PMID...clinical applications of optogenetics." Current protocols in pharmacology 75.1 (2016): 11-19. PubMed PMID... -
Quantifying DNA? Here are Five DNA Quantification Methods to Consider
TypeBlog PostPublishedJuly 21, 2020, 1:15 p.m....wavelength that they maximally absorb. The spectrophotometer measures these absorbances using UV-transparent...can’t distinguish between DNA and RNA. See our protocol about using UV absorbance to quantify DNA Fluorescence...specific for double strand DNA (as compared to spectrophotometric methods that measure all nucleic acids). ...methods can’t tell you and you don’t need a spectrophotometer or a fluorometer to quantify DNA this way....Addgene blog Read more about molecular biology protocols and tips Find the best way to elute and store ...Reference Find plasmids for your research Browse protocols for molecular biology, cloning, and more ... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog PostPublishedMarch 14, 2019, 1:54 p.m.... purification protocol involving iodixanol gradients. The current purification protocols for isolating...accumulate inside the producer cells and purification protocols often describe a process that begins with cell...Since the development of the exo-AAV isolation protocol by Maguire and colleagues, other researchers have...intravitreal injection to transduce bipolar cells and photoreceptors (Wassmer et al., 2017), and exo-AAV8 expressing...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ... -
Plasmids 101: Gram Negative and Positive Bacteria
TypeBlog PostPublishedAug. 2, 2022, 1:15 p.m....Image credit: Mark Perkins https://www.flickr.com/photos/occbio/7092385559 For more specific procedural...gramstain.html Smith, A. & Hussey, M. Gram Stain Protocols. American Society for Microbiology (2005). https...5c95a063-326b-4b2f-98ce-001de9a5ece3/gram-stain-protocol-2886.pdf Steward, K. Gram Positive vs Gram Negative...in marine biology and minoring in studio arts photography. References and resources References Silhavy...Hans-Christian-Gram.pdf Smith, A. & Hussey, M. Gram Stain Protocols. American Society for Microbiology (2005). https... -
OCaMP: A New Calcium Indicator for Neural Imaging
TypeBlog PostPublishedAug. 19, 2025, 1:15 p.m....for these lasers, with two-photon excitation at 1030 nm, enabling two-photon imaging without compromising...brightness, Less signal distortion from photoswitching or photobleaching Improved signal-to-noise ratio (SNR...current red and green GECIs aren’t ideal for two-photon imaging with newer lasers in the range of 1000–...ratio (Figure 3). Figure 3: One- and two-photon excitation and emission spectra of OCaMP. 1P excitation... -
Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME
TypeBlog PostPublishedJune 6, 2019, 12:41 p.m....Constructs using the photodimerizer system CRY2-CIB1 to regulate botulinum neurotoxin were also created....core of this tool is the endoprotease Botulinum Neurotoxin B (BoNT/B), which cleaves the synaptic protein... to SspB, brings together the split botulinum neurotoxin halves to enable VAMP2 cleavage, and suppresses...slices from mouse, and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids...gas-filled protein structure that water-bound photosynthetic organisms use for buoyancy. These proteins ... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m.... systems are still being developed. A detailed protocol for production of Gutless AdV can be found in ...looking at absorbance at 260nm using UV/Vis spectrophotometry. An absorbance of at 260nm corresponds to... Enjoy! References 1. Luo, Jinyong, et al. "A protocol for rapid generation of recombinant adenoviruses...adenoviruses using the AdEasy system." Nature protocols 2.5 (2007): 1236. PubMed PMID: 17546019. 2. He TC. et ...AdEasy System. 3. Jager, Lorenz, et al. "A rapid protocol for construction and production of high-capacity...high-capacity adenoviral vectors." Nature protocols 4.4 (2009): 547. PubMed PMID: 19373227. 4. Mittereder, Nanette... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog PostPublishedSept. 6, 2016, 2:30 p.m....particular the roles of proto-oncogenes, tumor suppressors, and their protein products. Proto-oncogenes consist...dividing out of control. One may compare the roles of proto-oncogenes and tumor suppressors to the accelerator...respectively. In the case of a mutation that turns a proto-oncogene permanently “on”, (now an oncogene), this...Downregulation of tumor suppressors and upregulation of proto-oncogenes can directly inhibit the ability of...H-Ras, K-Ras, and N-Ras) encoded by a family of proto-oncogenes. Ras proteins are special because they... -
Google Forums Round Up: First Impressions of NgAgo
TypeBlog PostPublishedAug. 4, 2016, 2:30 p.m....section asked basic questions about the NgAgo protocols used by respondents (transfection method, amount... to wait until someone else optimizes an NgAgo protocol for their favorite model system. The vast majority...system. I encourage you to share your results and protocols with the research community if you've had success...specifically set up a forum where NgAgo users can share protocols and tips Twitter - Tweet suggestions and tips ...lucky with NgAgo should explicitly share tips and protocol details with the scientific community on NgAgo...things (101 lessons on Spanish, C++, guitar, photography) and often dreams of a sunny day in Edinburgh... -
Participate in the Pets of Addgene Scavenger Hunt
TypeBlog PostPublishedApril 1, 2021, 1:15 p.m....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...