...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated...relies on a change in localization to the cell membrane and not a direct change in the fluorescent properties...

... streptavidin-coated magnetic beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and ...and target capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that...

...heads and long legs that protrude from the cell membrane, physically connecting the extracellular environment...the integrin, where it is resisted by the ligand embedded in the extracellular environment. This further...the extended open integrin conformation. During embryonic development, extended-open integrins enable tissue...

...hydroxybenzylidene imidazolinone (DFHBI). DFHBI is membrane permeable and non toxic to cells. Because it rapidly...split aptamer that can be used to investigate RNA assembly. The split aptamer is composed of a “Top” and ...fluorescent split aptamer for visualizing RNA–RNA assembly in vivo." ACS synthetic biology 6.9 (2017): 1710...

...proteins, which are directly trafficked to cell membranes. Membrane trafficking of farnesylated derivatives enables...fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered cyan). Without Cre recombination, RFP...

...optogenetic accuators to depolarize or hyperpolarize membranes. Traditional FRET reporters could not be used ...Venus fluorescent protein in conjunction with a membrane-inserted voltage sensing domain (LOTUS V; Luminescent...Universal Sensing of Voltage). With this sensor, membrane depolarization increases the BRET signal whereas...

... scientific mentor and choosing the right lab. Remember that your time is valuable! Don’t jump on the ...to plan out a very distinct and certain path, embracing uncertainty can help you both in your career and....org/10.1126/science.aba8039 Cordova FA (2016) Embrace uncertainty. Science 351:994–994 . https://doi....

...committee in graduate school. Thesis committee members provide expertise in designing and interpreting...to introduce yourself to and interact with the members of your field, and present your research. Face ...family. Those people are your support network, so remember to thank them verbally and often for their love...

...SDS-PAGE, and are subsequently transferred to a membrane. A primary antibody recognizing the protein of...is added, and will bind to that protein on the membrane. Then, addition of a secondary antibody allows...bind to proteins on the cell surface, or the cell membrane can be permeabilized to allow antibodies to bind...

...speed at which a confocal image can be acquired. Assembling an image pixel by pixel is slow; therefore, certain...detector such as a scientific CMOS camera. If plasma membrane dynamics are the main focus of the experiment,...dynamic samples Ex: 3D cell cultures, small model embryos >15 μm Thsee are some of the most challenging samples...

... in CRISPR–Cas9. The EMBO Journal, 37(10), e96941. https://doi.org/10.15252/embj.201796941 Pattanayak,...CTD and the entire REC lobe—do not structurally resemble any other known proteins; they are completely ...

...1), which may be easier to conceptualize. Just remember to switch them back before reporting the data!...use it! Tempting as it may be to skip this step, remember we here at Addgene frown upon statistical laxness...11237753. Primer design resources PrimerBlastDINAmelMfoldBLASTEnsembl More resources on the Addgene blog...

...single stranded, negative sense RNA virus. It’s a member of the Paramyxoviridae family of viruses, which...staining for alkaline phosphatase, expression of embryonic proteins, and promoter demethylation and expression.... Cells can be differentiated in vitro to form embryoid bodies which are compact balls of loosely organized...

... Biology 22:169–174 . https://doi.org/10.1016/j.chembiol.2014.12.011 Polstein LR, Gersbach CA (2015) A...Chem Biol 11:198–200 . https://doi.org/10.1038/nchembio.1753 Richter F, Fonfara I, Bouazza B, Schumacher...Chem Biol 13:443–448 . https://doi.org/10.1021/acschembio.7b00603 Additional Resources “Optogenetics Meets...

...are easily inserted using BbsI and Golden Gate Assembly. Either plasmid can be also linearized with XbaI...elements, like a repair template, using Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria...Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth Biol 7...

...It's December, and that means it's officially the start of Addgene's winter holiday celebrations! We'...with Addgene writers High Complexity Golden Gate Assembly with NEB's DADExperts: Matthew Norton and NEBGCE4All...

... former Co-director of Science in the News, and member of the SITN podcast team.     Amy Gilson recently...former Co-director of Science in the News, and is a member of the SITN podcast team. You can follow Amy on...

...Mark your calendars for November 16-20, 2020 for #DepositorWeek on Twitter!  That’s today through Friday...presented in this blog post includes data until November 9, 2020. Between then and the posting of the blog...

... that everything will ship out in time - from assembling the boxes, to inoculating tubes, and to packing...pic.twitter.com/eJbI0lLiGQ — Dr. Joanne Kamens (@JKamens) September 13, 2018 This massive scale reagent sharing led...

... the vectors are also likely to work in animal embryos based on earlier evidence showing that a CRISPR...PMID: 24954249 Find the Mulitplex CRISPR/Cas9 Assembly System Kit at Addgene here. Other Popular CRISPR...

... podcast series, we chat with new Addgene Board Member, Michael Koeris. Dr. Koeris did his graduate work...Resources on the Addgene Website Check Out the Other Members of our Board of Directors Become a Part of the ...

...intellectual property from our Tech Transfer team members Tony Cova and Chiara DeNatale and programming with...great European photo slideshow from our UK team member Benoit Giquel. Bluegene’s globetrotting impressed...

...micrograms of starting material; Gibson assembly or Golden gate assembly use a more direct approach, allowing...

...are coming along at a fast and furious pace. In December, as we've blogged about before, there was the ...via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014 January 30. (Pubmed) ...

...genome editing plasmids String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR), developed...

...how it's used. Chapter 12: Darrell's goals as a member of the Addgene board of directors - increasing ...stick to our core mission. Chapter 13: Outro - remember to step back and ask, "Why did I start doing this...

...to specific drugs in specific contexts, so he assembled a plasmid kit designed to do exactly that. As ... described in the journal Science Signaling in December 2014, he and his colleagues at Duke University...

...immunotherapy targets but are embryonic lethal, hindering studies post embryogenesis. Using C-CHIME and S-CHIME...

... former Co-director of Science in the News, and member of the SITN podcast team. You can follow Vini on...former Co-director of Science in the News, and is a member of the SITN podcast team. You can follow Amy on...

...algae cell is just the cell wall and cytoplasmic membrane. Vampirovibrio chlorellavorus will not grown in...appropriately named as spook, spookier, phantom, disembodied, shadow, and shade. Tongue-eating louse Cymothoa...

...create Phage Directory in a whirlwind three days in November 2017. #Phage researchers! I am working with...Strathdee, 🗡️Superbug Slayer 🗡️ (@chngin_the_wrld) November 8, 2017 Facilitating the search of phages for...

...benefits for both PI and trainee. DePace and her lab members have noticed that the meetings make the lab run...the PI has an opportunity to check in with lab members, determine how to work through obstacles, and take...

...all of the Addgene employees, the Addgene board members, vendors, Addgene supporters, and of course, Executive...integral part in its success. The Addgene board members were also recognized for their essential advice...

...filled with family, food, and travel, and you can’t remember where you put your flow cytometry protocol. Or...Tip 3: Share protocols and spreadsheets with lab members Now that you’ve organized your protocols, wouldn...

...participate in a brief fifteen minute interview. I remember leaving the interview hopeful to continue on in...the beginning, I spent a lot of time shadowing members of my team to learn what I was to do for my time...

...Addgene’s Lianna Swanson, who has been working with members of Davidson's lab to coordinate the impressively...F-Actin to mitochondria, from the nucleus to cell membrane, this collection has fluorescently tagged versions...

...for the first time, being introduced to all the members around the office one by one, I knew that I belonged...right concentration of a reagent during our HiFi assembly but decided to go ahead anyway. We were running...

...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended...Llamas Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids...

...came to visit us and watch fellow Run4Rare team members complete the Boston Marathon, he wanted to run ..., the R206H mutation makes ACVR1 responsive to members of the ubiquitous activin ligand family, including...

...insights they glean from a reading session. To remember what happened when drug X was added to cell line...with a specific gene knock out, often you have to remember the experiment, not the paper. To get around reading...

...performed with a probe surrounded by a semipermeable membrane, which is inserted into the region of the brain...passive diffusion of DA across the semipermeable membrane from the extracellular fluid into the perfused...

...destroy the invader. Wikipedia, accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/...(12): 5429–33. Photo: Wikipedia, accessed 25 November 2013. Author: James Atmos. http://en.wikipedia.org...

...Ever wondered where human embryonic kidney (HEK) cells originated from? Or why they are such a commonly...cell line model for cancer-specific research. Membrane receptors and ion channels are often exogenously...

... your research, check out this post! R Bodies: Membrane-Rupturing Microscopic Tools In this post, Jessica...also showed that R-bodies can rupture E. coli membranes. Read the full post for potential applications...

...These convert the amino acid tyrosine to betanin, a member of a class of pigments primarily found in plants...skill that takes practice and is difficult for new members of the lab or collaborators unfamiliar with these...

...the stochasticity of molecular interactions and embedding individual diversity to get from gamete to a visibly...replication and gene transcription during vertebrate embryogenesis. My interests, though, have proven unruly. Instead...

...useful for monitoring mitophagy. It’s important to remember that because Keima’s fluorescent relies on lysosomal...phosphatidylethanolamine (PE) and recruited to autophagasomal membranes, which leads to its degradation by autophagy. ...

... AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral vectors ...supernatant, the viral particles associate with the cell membrane and are taken into the cell via endocytosis. In...

...overhangs to your insert of interest for Gibson assembly. The steps following primer design and the PCR...to ligate our cDNA into the recipient plasmid. Remember to insert your DNA in the correct orientation ...

...make sure to rinse off the surface of the filter membrane with the surrounding solution to ensure excess...excess viral particles do not remain adhered to the membrane. Be careful when diluting - if you end up concentrating...