We narrowed to 868 results for: TOR

... post was contributed by Laura Desrochers, a postdoctoral scientist at AstraZeneca. Why I chose an industry...time working on something extremely risky or exploratory. Getting a job afterwards: Fairly or unfairly...an industry setting. Scientific community and mentorship: As an industry postdoc, you might find yourself...project’s or a team’s, it is possible that your mentor (and even their department) could be laid off while...Desrochers! Laura Desrochers is currently a Postdoctoral Scientist at AstraZeneca. She has a background...blog posts here Find 9 tips for a successful postdoctoral experience Read Joanne Kamen's 8 tactics for...

...My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, ... a heated stir plate to dissolve and requires monitoring so that the solution doesn’t get too hot because...approach gives me a bit of flexibility since I can store the cell lysates at -80 °C for up to 6 months before... completed in one long day, or the virus can be stored at 4 °C and buffer exchanged the next day. Refer...Production of recombinant adeno-associated viral vectors for in vitro and in vivo use. Current protocols...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

...localization, dynamics, and behavior of a POI. As an investigator, you want a fusion tag to be inert, to not produce...has gone into making FPs monomeric, yet many investigators remain ignorant of FP dimerization. Equally ... protein fusions Transmembrane proteins, i.e. receptors and transporters, are integrated into lipid bilayers...spiderweb-like pattern of tubules can be grossly distorted into dense stacked membranes termed Organized ...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum and live...

...production is Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins...using a desalting column or an ultrafiltration concentrator. As the name suggests, you may also concentrate... to their parental hybridomas. Aggregation and storage Aggregation of antibodies may impact their quality...testing. An aliquot of our recombinant antibodies is stored at 37 °C for a period of two weeks. After this ...we are also cautious during the aliquoting and storage process to prevent dropped tubes. Now that your...Varley P, Rockberg J, Chotteau V. Small-scale bioreactor supports high density HEK293 cell perfusion culture...

... (NHEJ) and alternative end joining (alt-EJ). Inhibitors to essential components of both pathways have...events (Yang et al., Int J Mol Sci). Adding these inhibitors to your culture media prior to and after Cas9...introduction increases HDR, just make sure to remove the inhibitors within a day or two to not affect the viability... one step further, you can inhibit cell cycle regulatory proteins responsible for the transition out of...to fuse an HR protein to the nuclease. Early HR factors fused to Cas9 have been shown to increase HDR frequency...committing a break to performing HR before other factors were loaded onto the break, potentially shuttling...

...small-molecule inhibitors of a particular PPI. In such experiments, small-molecule inhibitors are tested ...because FRET depends on a number of hard-to-achieve factors. For example, FRET requires that fusion proteins...signaling, integrin signaling, and transcription factor interactions (Kerppola, 2006). One of the first...between basic leucine zipper and Rel transcription factors (Hu et al., 2002). This would be more difficult...library to screen PPIs among 450 transcription factors in live fruit flies. Although BiFC open-reading...

...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An...This technique utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which result ... potential complications, there are additional factors to take into account with these types of experiments...and purification, which will require additional laboratory reagents, equipment, and expertise. Final thoughts...production of the Cas9 protein and a gRNA in your laboratory, once these have been acquired, genomic editing...Central PMCID: PMC4289409. 3. Rouet, Romain, et al. "Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease ...

...Our story today starts with a bit of Addgene history... As you may be aware, Melina Fan, her brother ...get Addgene from a concept to an up and running repository. As they built the foundation for what Addgene...previous years, this event was held at Executive Director Joanne Kamen’s home- a testament to the familial... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...Veckerelli is the Administrative and Operations Coordinator at Addgene. In addition to her operational responsibilities...

...yourself needing to brainstorm with a team. One major challenge with these brainstorming sessions is avoiding...with unlimited time? Another strategy for your brainstorm session is to start with something completely... ideas that I have found helpful in my team brainstorming sessions.  Allow the group time to think about...This is why perfectionists tend to also be procrastinators.  If you fall into this category, get out of...our lab space, our protocols and even our Lab Inventory Management system. I hope these tips help you ...

...bacterial world produce an assortment toxins to claim territory or kill competing microorganisms, but did you ...as a counterselectable marker in the destination vector. Successful cloning occurs when the ccdB gene is...unmarked genetic modifications in the yeast Pichia pastoris. By combining both positive and counter selection...induced by methanol. Once linearized, the delivery vector can be integrated into the genome to introduce .../sok toxin-antitoxin system, they were able to monitor lacZ activity over 48 hours in vitro without antibiotic...marker for unmarked genetic modification of Pichia pastoris." FEMS yeast research 9.4 (2009): 600-609. PubMed...

...lysozyme to reduce basal expression levels; expression vector cannot have p15A origin of replication F- ompT...higher T7 lysozyme expression than pLysS; expression vector cannot have p15A origin of replication F- ompT...codons not normally used in E. coli. Expression vector cannot have p15A origin of replication F- ompT ...alleviates inclusion body formation. Expression vector cannot have p15A origin of replication fhuA2 ...Cis-repression of the E. coli T5 promoter (found on vectors such as pQE or similar), inducible under IPTG (...lac repressor on the pREP4 plasmid). Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15...

...signaling pathways. Rho guanine-nucleotide exchange factors (GEFs) activate their partner Rho GTPase, leading...to reveal even more insights. Super-duper lipid editor And if recruiting proteins to the cell membrane...phosphatidic acid signaling (such as Hippo, AMPK and mTOR pathways) and to introduce unnatural head groups...membrane or lysosomes and its activity can be monitored by phosphatidic acid probe GFP-PASS. B) Time series...Additional resources on the Addgene blog Viral Vectors 101: Optogenetics Additional resources on Addgene.org...Activity-based directed evolution of a membrane editor in mammalian cells. Nature Chemistry, 15(7), 7....

...know is your area code. Senators: To find contact information for your senators all you need to do is select...active research scientists (scientists who hold a doctorate degree, are currently working full-time, and have...edition. Cambridge university press, 1988. AAS: Historical Trends in Federal R&D NJ.com: Rush Holt: Lawmakers...Scott Pruitt says carbon dioxide is not 'primary contributor' to global warming The New York Times: With Trump...affected by Trump’s immigration ban Studies in History and Philosophy of Science Part: Democratic values...

...list from your current P.I., lab manager, or administrator containing all of the materials that you’ve ...financial values. All too often, we see early investigators making unnecessary short term sacrifices that...negotiations can still go horribly awry when several factors aren’t taken into consideration. 1. Always focus...solutions to reach your goals. For example, a junior investigator turned down a faculty position because none ...Happy Sciencing, Damien   Damien Wilpitz is a laboratory research management consultant and coach in Boston...research scientists create productive and manageable laboratory environments. His business skills enhance his...

...construct (about 8 in Brainbow-1.0 mice.) The combinatorial effect of multiple, independent recombination...woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), commonly used to increase transgene...Addgene also has two Brainbow adeno-associated viral vectors (AAV) available - AAV-EF1a-BbChT and AAV-EF1a-BbTagBY...of neural circuits, such as connections between motor neurons and the neuromuscular junction. In zebrafish...plasmids References: Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous...PubMed. Multiplex cell and lineage tracking with combinatorial labels. Loulier K, Barry R, Mahou P, Le Franc...

...combining AcrIIA4, a Streptococcus pyogenes Cas9 inhibitor, with the LOV2 photosensor from Avena sativa, ...-CRISPR plasmids Tagging human transcription factors with GFP Kevin White’s lab recently deposited a... of CRISPR plasmids to tag human transcription factors with GFP. As part of the Encyclopedia of DNA Elements...plasmids will help researchers map transcription factor localization and binding via ChiP-seq. Learn ...Learn more about ENCODE Find the transcription factor tagging plasmids If you have a new CRISPR tool you...

...Addgene with our Executive Director, Joanne Kamens. As a regular contributor of careers blog articles, ... our visitors made sure to tell us what we already knew about our Shipping Operations Coordinator: “Kory...teams at Addgene. An introduction to plasmids with Director of Biology Lianna Swanson was very popular, with...

... take into account chromatin context and other factors, they are easy to use and can reduce off-targets...nicking/cleaving of DNA occurs, not for deadCas activator/repressor experiments where only localization ...- GUIDE-Seq and integrase-defective lentiviral vectors - which rely on insertion of DNA by the NHEJ pathway...RNA-seq would be an appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would... (e.g. in a gene in a similar pathway, a growth factor, etc.) this control is usually not very meaningful...and off-target risks will vary depending on many factors. Hopefully now you have all the resources you need...

...and luckily genetics is a sector which has been rapidly growing. These factors left me optimistic that ...and the pro’s and con’s of working in different sectors. Luckily, I have worked for a variety of different...options for going into parasitology but another major factor held me back; I do not have a second language. ...allowed me to try out working in many different sectors and to discover for myself what potential job opportunities... Are Open at Addgene Learn How to Choose a Good Mentor for Your Professional Development ...

...signaling activity and cell phenotype. One method for monitoring live single-cell kinase activity is FRET, but... distance and the orientation of the donor and acceptor proteins, which can be difficult to predict. ...disadvantages when compared to FRET reporters. First, KTRs monitor global kinase activity that occurs in the cytoplasm...experiments. Third, KTRs are not well suited for monitoring kinase activity that occurs during nuclear envelope...the nucleus, there’s no nuclear localization to monitor). Whether you’re looking for a ready-to-use KTRs...