We narrowed to 877 results for: HA

... transcriptional read-through. Chris Voigt’s lab has characterized a set of prokaryotic terminators and...for eukaryotic termination are understood and it has been noted that each eukaryotic RNA polymerase terminates...combination with certain viral packaging systems has been associated with reduced viral titer, but improved...Expression Levels of Lentivirus-Delivered Transgenes but Has the Potential to Reduce Viral Titer in a Promoter-Dependent...

...to see if anyone has published a protocol that would work for your system. Addgene has depositor submitted...Sep 30, 2020 by Marcy Patrick. CRISPR technology has been widely adopted for genome editing purposes because...Mammalian CRISPR expression systems Each model system has its own best practices for efficient delivery of ...

...understands (synthetic biology), building a product no one has ever seen before (flowers that change color throughout...but the experience is invaluable. This opportunity has given us the chance to develop the technology we ...the business training to make the most of it. It has also allowed us to really analyze what we want out...Nikolai Braun grew up outside of Washington D.C. and has always enjoyed exploring the outdoors and experiencing...

...single-cell level. CRISPR libraries CRISPR screening has become a popular tool for probing cellular pathways...than knockout screens. The Jonathan Weissman lab has deposited genome-wide CRISPR activation (CRISPRa)...to browse. Our Guide to Plasmid Pooled Libraries has tips on using these libraries and designing screens...

...channelrhodopsins. HcKCR1 has a maximum absorption of 540 nm and two-photon excitation around 1,040 nm, has larger photocurrents...anti-c-Myc [9E10] (Evan, et al., 1985), the Myc tag has become one of the most commonly used protein tags...

...require heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating...showed that the receptor-based fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high...recent months: Our selection of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use...

... development, and response to environment - ABRC has kits for all these topics and more. Downloadable ...ordering account. Using Arabidopsis in the classroom has many benefits. Engaging students in active learning...working with animals. Because Arabidopsis is small, has a short life cycle, and is relatively easy to grow...

...KSgenome and seeing the intricacies of DNA repair has emphasized how essential these studies are to improving...alternative non-homologous end joining (Alt-NHEJ), has been harnessed for CRISPR knock-in applications. ...29266098 Creating knock-in mouse models with CRISPR has been challenging, with protocols using dsDNA templates...

...for mapping where a protein binds genome-wide, and has been widely applied in many model organisms and cell...proteins (Figure 1). Importantly, since Protein A has weaker affinity for mouse IgG, it is helpful to incubate...prior to the addition of Protein A MNase. Once MNase has been localized to target sites, the nuclease is briefly...

...Show that as the story has progressed, you or your lab or science or your CV has grown as well. Find your...Task, Action, Result) interview technique, which has been addressed in much better pieces than I’m able...

...measuring DA and other neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid... dLight1 dopamine sensor In 2018, Lin Tian’s lab has developed dLight1, a molecular tool to detect and...here! The GRABDA dopamine sensor Yulong Li’s lab has created the GRABDA sensor based on a similar strategy...

...AAV5  pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) 121538  AAV5  pOTTC1484 - pAAV SYN1 HA-hM4D(Gi) 119741  AAV5 ...iGABASnFR GABA sensor (pAAV.hSyn-FLEX.iGABASnFR) has good membrane localization and brightness. The F102G...

...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended... antibody libraries to generate new sdAbs. There has also been some interesting work done developing transgenic...al., 2021.   The single-domain antibody field has come a long way since the serendipitous discovery...

...inhibitory GABA-ergic interneurons, Gordon Fishell’s lab has developed a new AAV approach using regulatory elements...fluorescent protein - mScarlet! The Dorus Gadella lab has recently developed a synthetic red fluorescent protein... imaging and in protein fusions. The Gadella lab has deposited all mScarlet constructs with Addgene. ...efficiency Blue flame depositor Yannick Doyon’s lab has developed a new method to increase CRISPR editing...increases the probability that the locus of interest has also been edited! Thus, ouabain selection enriches...coselection are available at Addgene, and the Doyon lab has also provided a detailed protocol at Protocol Exchange...Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many applications in biotech...

...and insert a linear fragment of DNA (insert) that has been cut by compatible restriction enzymes. An enzyme...one found on the left side of the image below. It has a promoter (blue arrow) followed by the restriction...finished plasmid   Once it looks like your ligation has worked, you will need to pick individual bacterial...purpose. Ideally, once you know that your plasmid has an appropriately sized insert, you should send it...plasmid for more details. Once your complete plasmid has been verified, you’re ready to get experimenting!...

...! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000 times (over 800 orders...broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin ...

...In the last decade, the use of degron tags has become increasingly popular for the modulation of endogenously...protein levels can return to normal once the drug has been removed. The degradation and reversal are rapid... its first use as a protein regulation tool, AID has gone through an additional round of evolution (AID...the system, but the general concept of the system has remained the same. An obvious drawback of the system...

...FIQ or mStayGold (Ando et al., 2023). This variant has similar photostability and brightness compared to... of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with slightly...purify or image a recombinant protein, the FLAG tag has proven itself a good option for essentially all applications...western blots, though the original parent clone (M2) has been widely used in a variety of applications. Find...

... disrupting normal brain function. Chemogenetics has become a popular tool in the field as it provides...chemogenetics are instead that it is noninvasive and has longer-lasting effects. Building your chemogenetic... agonist for muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have...are already approved for clinical use. Varenicline has been further modified to create ultrapotent PSEMS...

... ddPCR.) qPCR Quantitative/real-time PCR (qPCR) has been used for copy number assays for years. qPCR ...and quencher to quantify specific amplicons.  Sybr has high background noise, due to the large amount of...AT-rich DNA and the length of the amplicon and it has a higher rate of false positives (Colborn, 2008; ... used to accurately call copy numbers, though it has been shown to identify trends correctly (Muhamad,...

...Not the case, as I’ve discovered.) As someone who has now seen both sides of the postdoc experience, I’... lab is closely involved with clinical trials or has ties to industry, the drug discovery process can ...Genentech, will not. The point is that each company has a unique approach, and some might suit your needs...currently a Postdoctoral Scientist at AstraZeneca. She has a background in cell and developmental biology and...

...the growing spectrum of fluorescent proteins (FPs) has been an invaluable resource for studying the organization...sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists to tag and light up endogenous..., and T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The...characteristics. For example, Masato Kanemaki's lab has developed a CRISPR/Cas-based system for tagging endogenous...

...community has access to all of the latest resources? In response to this challenge, Addgene has launched... why recent work in the field of cancer research has focused on picking apart the complex cellular pathways...at Vanderbilt University in Nashville, Tennessee has already demonstrated that a number of novel small...

...host cells immediately prior to their transduction has been shown to increase AAV transduction efficiency... such cationic reagent that is commonly used and has been shown to increase transduction efficiency by...single-use aliquots. Never use a polybrene solution that has been thawed more than once. Fibronectin is another...polybrene (e.g., hematopoietic or primary cells), and has been shown to increase transduction efficiency by...

...regulatory elements. To solve this problem, the Zhang lab has previously used SaCas9 (1053 aa/3.2 kb) with AAV ... these options (2.7 kb with 24 nt guides), so it has the potential to eclipse any of the CRISPR nucleases...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...be the best option for HDR, but this possibility has not been experimentally confirmed. Researchers have...

...for the presence of an insert. Traditionally this has been done with restriction enzyme digest; however... Find Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized in...insert-specific primers but also tell you whether the insert has the correct directionality. Verifying the insert ...and are more likely to work in a PCR reaction that has bacterial debris. Use a positive control. A good...

...Fluorescence is the emission of light by a substance that has absorbed light. The emitted light is at a longer ...protein Excitation & Emission (ex/em): Each FP has its unique ex/em peak. Therefore, choose FPs that...detect the emission. For example, if your microscope has only two lasers, at 488nm and 561nm, you will not...Some proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be taken into...

...10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through a 0.22 µm...

...process, though the process has changed a lot over the last twenty years! This has helped us develop a fast... request going forward.  After another scientist has placed a request for your plasmid, they must complete...

...Davis School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...mission This is the first NIH grant that Addgene has received. Addgene prides itself on being financially...The timing is ideal as the NIH’s BRAIN Initiative has been funding laboratories, such as Dr. Trimmer’s ...

...attributes (which turned into www.fpvis.org).  That page has been very useful over the years, and when researchers...system that allows users to see when a given page has unmoderated changes and to view previous versions... online collections of FP spectra. Every protein has a page with all the information in the database relating...

... dronpa fusions. With these various designs, Xin has been able to develop photoswitchable Intersectin,...Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs in a variety... of cells that actively produce the HMOs. Fatima has demonstrated the successful production of both types...

...AAV2 pAAV-CaMKIIa-HA-hM4D(Gi)-IRES-mCitrine 121538  AAV5 pOTTC1484 - pAAV SYN1 HA-hM4D(Gi) 121539 ...121539  AAV5 pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) New chemogenetics AAV Plasmid Serotype Name 119741  ...

...birth several weeks previously. While this claim has yet to be independently verified, the watching world...Kingdom’s Nuffield Council. While the Nuffield Council has recently published a report on the social and ethical...considerations, in making sure that the research has been adequately proven to be reasonably safe and ...

... to the original paper that everyone in your lab has been referencing for the last 10 years, and everything...communication further up the collective priority list. There has been progress—for example, a number of funders (e.g...joined Elsevier after her postdoctoral studies, and has held various editorial roles on review and primary...

...easy. Most of our equipment has been donated by local companies and BosLab has grown into an organization... biologist with a Ph.D. from UNC-Chapel Hill and has extensive experience targeting RNA with small molecules...

...amplified and none are lost. Each Addgene library has its own depositor transformation and amplification...product for next-generation sequencing Once the DNA has been extracted, the representation of the library... success of your amplification. I hope this post has helped clarify some of the important considerations...

...derived from the same cell type; however, their usage has been limited by technical problems. For instance,...binary systems in which transient protein expression has a long-lasting effect, such as Cre- or Cas9-mediated...effects. Third, protein purification (once a protocol has been established) may be less laborious than viral...

...sequence if necessary. The example plasmid on the right has a total size of 7.3kb, including a 1.2 kb insert....resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, with the ...reference point so you can easily assess how far the gel has run. Bonus: The dyes also run at predicted sizes ...

...Griesbeck of the Max Planck Institute for Neurobiology has been working on genetically encoded indicators of...imaging neurons, Griesbeck adds that this color change has a particular advantage for observing events taking...Maarten Merkx of Technische Universiteit Eindhoven has similar interest in FRET-based sensor proteins, particularly...