...the conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling...and more photostable than fluorescent proteins, making them ideal for single-molecule fluorescence techniques...deposited an array of constructs for tagging the kinesins KIF5A and KIF5B as well as backbones for generating... will be useful not only for future studies of kinesins but for many other proteins of interest too! Find....A.S., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation...voltage imaging. A) ASAP5 selected for its improved kinetics and responsivity in screening ASAP3 variants. ...different illumination regimes with fast activation kinetics and high responsivity near the resting membrane...

...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...undergrads back in grad school. Both were hard-working scientists-in-training who got the job done, but...left him/her with small tasks and s/he was always asking for more work (this is a big hint that you are ...on what you should be doing : writing a grant, thinking about the next project, reaching out to other ...really helpful. “What is the best/worst part of working with X?”  is going to get you what you really want...grateful for the feedback, but at least you can start making changes for the best. Positive feedback is equally...Scientists Series What Makes a Good Mentor? Tips for Working in Industry and Success in Management Resources...

...researchers learn many transferable skills while working in the lab and by offering advice on areas where... jobs needed to be done (Who is responsible for making the running buffer this week?). Scientists negotiate...approached it as a creative problem-solving exercise? Asking questions like the below can help you understand...you are in the middle of a frustrating situation. Asking for opinions from a neutral third party can be ...offer conflicts with a pre-planned vacation I’m looking forward to. How can I negotiate my starting date... a tenured professor or signing contracts when working for a business development team, negotiation is...strengthen your relationships with others who know that working with you will benefit them too. Ed's note: Salary...

...structural biology, Byron Rubin, a crystallographer working with Jane Richardson, provided an initial solution...model. Until then, you can only wonder what I am talking about. How do researchers use physical models of... over and over in their hands. They then start talking to themselves as they compare the features displayed...magical teaching moments. The models function as thinking tools – eliciting questions like “what does this...investigated in their labs. Soon after we began working with educators in our Center for BioMolecular Modeling...complementary DNA strand. And this is why we love working with researchers on a model. The little knowledge...Jennifer Doudna’s new book A Crack in Creation. Making models more useful through flexibility It is common...

...of His-tagged human kinases in bactera. Listen to Our Segment on The Human Kinase Domain Constructs Kit...We’re breaking into more audio and video on the Addgene Blog and Addgene website. As we push forward ...CRISPR X from Michael Bassik's Lab - A new tool for making diverse, localized point mutations. Listen to Our...Segment on Magneto 2.0 Hot Plasmid 3: The Human Kinase Domain Constructs Kit from John Chodera's Lab -...with Eye Researcher Connie Cepko Get Advice on Picking a Graduate Lab and Project Check Out 5 Science ...

...scientific discovery and career development. They are working with limited resources but limitless potential....by facilitating connections to mentorship and networking opportunities, publishing assistance, and more...international scientific community and are always taking steps to increase plasmid distribution and disseminate...distribution As you can see from the chart below, we are making great progress toward spreading the Addgene community...countries. As part of these efforts, we will be working with Seeding Labs to award plasmids to research...well as shine a light on some of the great minds working in developing countries. As the world continues...

...been extremely helpful for scientists that are looking for more detailed information about this new technology...topics you want to learn more about. Our post on “Making Your Own Competent Cells” has been extremely popular...additional questions. We’re happy to see scientists making their own cells to save money and create the tools...feature in the coming year. Finally, we are always looking for guest bloggers who want to share their research...Replication By Kendall Morgan Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids...Marcy Patrick Will You Be My Mentor? Finding and Asking for Mentoring Support By Joanne Kamens Plasmids...

... you’re an oldbe trainee!) can be pretty nerve-wracking. So how can you be the best trainee possible? ...articles on building successful mentor relationships, asking for mentor support, and choosing your mentor. What...And I mean everything! The bare minimum should be taking notes on general protocol steps and other tips ...the moment….but you’ll probably forget and end up asking your mentor later if you don’t write it down. And... to your mentor. Pro tip! Consider physically walking through the protocol, by going to each space in...tell a trainee is understanding and things are ’clicking’ for them. It’s often easiest to train individuals..., What Now? Will You Be My Mentor? Finding and Asking for Mentoring Support   ...

...Mortensen, a PhD candidate at Northeastern University. Working with plants doesn’t always have to be a time-consuming...recently overcome this problem in C. roseus by working with young seedlings and optimizing several other...transient leaf transformation: Check if there are working protocols available for your species This will ... your tissue. Run a control with Agrobacterium lacking the GUS gene since plants can have endogenous GUS-like.... Successful infiltration can often be seen by sinking of the tissue (see Figure 3). This method is probably...GV3101(pMP90), which works well for us. Use a well working vector system This allows you to quickly assemble... you started. Once you have established a well working protocol in your lab, it can become a strong tool...

...the clock for others to dive in. The real cost of making data accessible A more reasonable concern with ... the scientific community is thus not averse to making data available, but there remains concern about... by others. Other new data initiatives are also taking root. Nature Publishing Group recently launched...efficient searching and extraction of information. Looking a little further ahead, journals may move towards...Instead of providing a “dead end” figure or table, clicking on the published chart could reveal the underlying...reader as possible. Rather than adding to demands, making effective data sharing the norm should open up ...Canada where he studies protein-serine/threonine kinases implicated in human diseases. His spare time is...

...epigenetic silencer Recombinant antibodies for marking astrocytes MagIC-Cryo-EM for sample enrichment...minimizes sample loss while also supporting imaging, making it easy to identify dozens or hundreds of particles...molar ratio) with 10 minutes of glutaraldehyde crosslinking: representative micrograph (C), particle orientation...could be overcome with methods such as chemical crosslinking or tilted-stage imaging. A truncated form of...compatible with the buffer additive MgCl2 and the crosslinking agent glutaraldehyde. Adding RvLEAMshort to ...j.cell.2021.03.025.    Recombinant antibodies for marking cell types in the nervous system By Ashley Waldron...filament expressed in astrocytes throughout the brain, making it a popular target for labeling these cells. Check...

... pathway. Those can include cytokines and chemokines; enzymes like kinases, phosphatases, and ubiquitin...false positive signal. Dump gating When you are working with heterogeneous cell mixes, like bulk lympho... buffer solutions. Fixing causes protein cross-linking, killing the cell but preserving the cellular state...gating uses the same color for different markers, making it easy to gate on the negative cell population...investigating negative regulators of T cells and cytokine signaling in tumor immunity. When he's not in ...

...of naked DNA. Other investigators who had been working to develop high efficiency gene targeting approaches...the addition of gene-specific 1 kb homology arms flanking a FLOXed neoR gene. pAAV-SEPT-Acceptor, the preferred... CRISPR-based approaches for the generation of knockins via homologous recombination with a homologous...application - the creation of sequence-specific knockins - for which the AAV approach is most well suited...suited, since it routinely generates knockin efficiencies of 1-40% with G418-selected clones. However, there...approach requires G418 selection of infected clones, making it unsuitable for most in vivo applications.  Third... CRISPR-based approaches for the generation of knockins, it is not always ideal. Because of these limitations...

...in vitro kinase reactions. Often, however, researchers lack knowledge of the regulatory kinase, access ...access to active kinase for the reaction, or have difficulties with kinase specificity in vitro. The establishment...phosphorylation sites. If knowledge of a candidate regulatory kinase is known, it’s possible to generate non-phosphorylated...phosphoserine into recombinant proteins, such as the kinase MEK1 (5). MEK1 is inactive without requisite dual...create a pSer/phosphomimetic dual mutant) elevated kinase activity over 70-fold over dual phosphomimetic ...

...every day, sharing ideas for the company, but also talking about sports, movies, and just about anything. ...cite lunch as one of their favorite parts about working at Addgene. We’re a small non-profit, so we can...Chef-style competitions Every few months, we’ll have a cooking competition featuring a selected ingredient. Anyone...cook-off last week, and the featured ingredient was pumpkin. Everyone gets to vote for their favorites, and...the stiff competition or says something about my cooking.   This is a creative (and delicious) team building...work better and keeps us smiling even when we’re packing the 400th plasmid that day.   As we head into Thanksgiving...

...Does this sound familiar? Well, when it comes to working with Addgene, we certainly hope not! One of our...email. You need help right at the moment you’re looking for it, or you may just prefer to troubleshoot ...icon, search for answers, and choose a result. By clicking on the result links themselves right from within...ll navigate to articles answering your questions making it easier than ever to get the support you need...some reason you can’t find the answer that you’re looking for, or you just have a general question or concern...valuable feedback. This is Addgene after all, and making your research easier always comes first. Resources...

...fused to calmodulin (CaM) and the chicken myosin kinase calmodulin binding region RS20. When calcium is...place for the biosensor to be expressed. Are you looking at an intracellular or extracellular molecule? ...extracellular glutamate sensor pCMV(MinDis).iGluSnFR. Picking your reporter Don’t forget about the other end ...with your lab and experimental setup? Consider what kind of instrumentation you have available, and the conditions...and multi-color imaging experiments. Finally, what kind of system will you use to detect your fluorescent...method depends on what type of information you are looking for. Below we have summarized some of the common...

...its value. My master’s strengthened my critical thinking and problem solving abilities, increased my technical...writer,” among others. Don’t discount job postings looking for PhD applicants since many will also accept ...people describe their actual work. To do this, try looking at LinkedIn profiles of people who have masters...Is their career path one you could see yourself taking? If so, consider setting up an informational interview...degree prepared them for their career.  2. Are you looking for career advancement and growth? A master’s is...project.  3. Do you prefer being a generalist? Working on a variety of projects and in different fields...

...equipment acquisition, public speaking and personnel management?  If you are thinking about a non-academic path...advisors learning how to be a scientist. It is shocking how little pre-work most PhD students and postdocs...learn how to be a scientist. Your PI should be the kind of mentor that will teach you how to: interpret ...with the potential advisor and don’t be shy about asking hard questions. Talk to as many lab members as ... time with everyone. Additionally, if you are looking for more resources to help you choose a good mentor...

...have an opportunity to improve my networking with mentors who are working in USA and Europe, something that...who migrated from their country some years ago looking for their career improvement) and bright students...world in flux. The professional development and networking resources offered by programs like IMP USA-Europe...Institute of Technology, Boston University, Johns Hopkins University, the Salk Institute, and the Karolinska...and contacts with Spanish laboratories to start working on projects together. List of discussion topics...Addgene Read Other Career Blog Posts Read the Networking for Scientists Series Read our Blog Post about...