...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...

...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...

...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...

...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...

...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...

...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...

... 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 ...

...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 ...

...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...

...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 ...

... 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...

...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...

...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...

...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...

...  Lowering cytotoxicity. Different envelope proteins have different levels of cytotoxicity and the mechanisms...pseudotype your viral vector, you use the same protocol for generating lentivirus, but you use a different...mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein causes cells... form syncytias that cause the cells to die. Cytotoxicity can also be cell type dependent: if a cell has... Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents for Pseudotyping Lentiviral Particles...

...yield of the donor (number of emitted photons per absorbed photons) and the extinction coefficient of the...microscopy (FLIM) method. Acceptor photobleaching method The acceptor photobleaching method is simple but limited...commonly used to measure FRET: the acceptor photobleaching method and the fluorescence lifetime imaging... the donor is quenched when FRET occurs. By photobleaching the acceptor, you release the donor's quenching...

...reaction. Below is a step-by-step overview of the protocol: Prepare ssDNA template: The wild-type plasmid...depth. Can I use my favorite plasmid with this protocol? Maybe. Not all plasmids will work for the one-pot... BbvCI site(s). If so, the nicking mutagenesis protocol will still work if the orientation of the BbvCI...multi-site saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis...Methods in Molecular Biology In Vitro Mutagenesis Protocols, pg. 103-109. PubMed PMID: 20676978. 3. Wrenbeck...Scale Using Deep Mutational Scanning.” Nature Protocols 9.9 (2014): 2267–2284. PubMed PMID: 25167058. ...

... not photostable, and were presented in the study to support the uniqueness of the photostability of the... and non toxic to cells. Because it rapidly photobleaches, DFHBI can only be used for qualitative, but... Corn and its small molecule makes Corn more photostable than Spinach and Broccoli, and useful for quantitative...increased brightness compared to Mango, and all are photostable and useful for both live and fixed cell imaging...Imaging RNA polymerase III transcription using a photostable RNA–fluorophore complex." Nature chemical biology...

...the target site, while Cas12a requires a PAM (Protospacer Adjacent Motif) for target cleavage. This test...40 min incubation. A scalable, easy-to-deploy, protocol for Cas13-based detection of SARS-CoV-2 genetic... Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller... acid detection with CRISPR nucleases. Nature Protocols 14:2986–3012 . https://doi.org/10.1038/s41596-...Wilson, M. Z. (2020). A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic...Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol for detection of COVID-19 using CRISPR diagnostics...

...production standpoint, it wasn’t ideal for our Lab Protocol videos. It’s really nice to have a scientist on-screen... on-screen, somebody who really knows the protocol and can guide viewers through. I know cameras, but ...thankfully volunteered, and we decided to focus on a protocol that didn’t require any lab shooting, just animation...setup for the lab, so we can get back to producing protocol videos?   Figure 4: The remote directing...the pandemic with some incredibly strict safety protocols to keep Addgenies safe. Space was at a premium...why not check out all of the written and video protocols at Addgene? If you need help in the lab, we have...