We narrowed to 890 results for: IND

...with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...endogenous ERK, and its localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates...Researchers commonly use rectal bacterial infection to induce tail swelling in C. elegans, but activation of ...cells by 24-48 hours exposure to blue light also induced tail swelling in ~75% of worms (see below figure...photoswitchable kinases in my experiments? You can find the plasmids from Zhou et al here. In addition, ...empty backbones in the Michael Davidson Collection Find Photoswitchable Fluorescent Proteins ...

...to autophagy-inducing conditions. Rosella makes it easier to study the mechanisms behind autophagy by... cells. Autophagy (Greek for “self-eating”) is induced by a lack of nutrients and targets cytosol and ... via proteasome degradation. Autophagy is then induced and quantified by measuring the amount of radioactivity...radioactivity in the culture supernatant (this is indicative of protein degradation). To control for protein...cytosol in an inactive form unless autophagy is induced. Then non-selective macroautophagy of the cytosol...some conditions it aggregates in an autophagy-independent manner. Additionally, tracking Atg8p or LC3 doesn...fluorescence. After 4 hours of nitrogen starvation to induce autophagy, red but not green fluorescence accumulates...

...channels, the team will be able to validate their findings in mammalian cells with both human and murine ...cytokines actually get out of keratinocytes and induce their effector functions. It’s an important process... Hospital Zurich. IL-36 cytokines are a potent inducer of an autoinflammatory loop: keratinocytes both...Lin got me interested in TS when I was a newly independent faculty member studying sex differences in thoracic...thoracic aortic disease,” Prakash says. In induced pluripotent stem cells derived endothelial cells from...thirty GNB1 missense variants identified from 58 individuals with GNB1 disorder since the identification of...allow us to enhance our biosensor repository as we find ourselves lacking several crucial biosensors that...

...version, but if you find yourself doing your own conjugation, it is important to keep in mind that you will ... be useful. Mainly because, well, antibodies are kind of hard to see. To solve this issue, researchers...antibody conjugates in an earlier post, where you can find some additional tips for working with these tools...affinity for avidin and streptavidin, both of which can bind up to four biotin molecules and can themselves be...with significant multiplexing potential. You will find antibody-oligonucleotide conjugates used in assays...protein-protein interactions with other peptides, like an Fc-binding domain (Figure 1).  Figure 1: Examples of...). If some of those lysines are in the antigen binding region or even just nearby, then conjugation could...

...skills, individuals also tend to have their own specialties. This type of team can exist indefinitely. If ... - you just want the necessary skills. You might find that your next paper is going to require help from...project that never ends is a poorly planned project indeed. Mission team A Mission Team is assembled to make...this year, I joined Addgene’s Green Team, a like-minded group of co-workers who want to make our organization...creating and strengthening those social bonds. So never mind how much better your work and productivity will ...someone to decide, someone to delegate, etc. If you find that there is not someone (or someones) taking on...easily discover you need to add some people if you find there’s a need for an expertise your current team...

..., coughing, etc. Also, a constant in-flow of individuals wherever the cell cultures are kept will increase...Contamination! Tissue culture media often have indicators included in the media that turns yellow from ...trigger a series of chemical reactions and this induces a noticeable color change. This method has high...fluorescent signals produced by these bacteria can hinder correct result interpretation. Also, the signals...Also, prolonged use of a particular antibiotic can induce resistance for other bacteria apart from mycoplasma... should be cleaned regularly and dampness of any kind should be avoided. Distilled water provided to maintain...because you are not the first person to face this kind of problem. Educate others when you can and help...

...that absorb photons for energy conversion or the induction of intra- or intercellular signaling pathways ...extra-chromosomally long-term expression and low ability to induce innate immune responses (Fenno et al., 2011). However...site-specific recombinases such as Cre or Flp that induce DNA recombination at defined recognition sites ...between mismatched recognition sites, its presence induces first opsin inversion, and then lox sites excision...opsin gene; in Cre expressing cells, however, Cre induces the recombination of the double-floxed opsin construct...FLEx-based vectors to study the neural circuitry behind the mechanisms of sleep and anesthesia. In Cre ...that it is possible to separate neuronal inputs to induce aversion- or reward-related behaviors. The high...

...preliminary data to establish your own lab and get an independent investigator grant (e.g. an R01 from the NIH,...makes time for you, as needed, this is a strong indicator that this person cares about your science and ...the reagents and outcomes. This is preferable to finding out you've been doing something all wrong or that... as they can result in spurious yet intriguing findings that fail to replicate. Plan experiments logically...great discoveries in science arose from surprising findings when studying a seemingly unrelated problem. However...As you get better at telling your story, you can find clearer ways to consolidate or present your data...you give your talk or poster to enough different kinds of audiences and if you pay attention to how the...

...standardized validation techniques for academia, industry, and especially the clinic. Popular validation...enhancing confidence and reproducibility of a study’s findings. Table 1: Biased Genotyping Options Technique... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...experiments Unbiased detection methods are excellent for finding evidence of DSBs throughout the genome. However...due to >3 bp mismatches or that are sequence-independent are rare, but they are detectable using just ... et al. "High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells." Nature...high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations using next-generation sequencing." BMC...

...the hopes and concerns of these communities. This kind of engagement and two-way dialogue is crucial if...meetings of experts and leaders from academia, industry, community, education and government; advising...make their own decisions about genetics, both as individual consumers of technologies and as societal participants...three years. pgEd is a small team comprised of individuals from diverse backgrounds and disciplines, and...is to liaise with different organizations and individuals, to identify ways of collaborating, and to organize...academic venues such as scientific, educational or industry conferences, as well as schools, community events...and in helping you decide whether you enjoy this kind of work. It is also beneficial to hone your non-...

...phosphopeptide binding domain FHA. The FHA1 domain binds to phosphorylated PKA substrate, inducing a conformational...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...undergoes a conformational change that permits it to bind to the CIB1 (cryptochrome-interacting basic-helix-loop-helix... by measuring growth of the ΔguaB pDCAF3 strain. Find the decaffeination plasmid here. Quandt et al. ...adapt similar FLINC biosensors to work in your lab. Find the FLINC biosensors here. Mo et al. Nat Methods... extinction coefficient when compared to sREACH. Find the FLIM-FRET acceptors here. See Addgene's FRET...

...allows for tagging and visualization of specific, individual proteins under endogenous regulatory control....your own gene-tagging constructs below. Read on to find out how these plasmids work and to discover more...cells so they can go to work. Figure 3: Inducing the double strand break and introducing the tag...BioRender.com. Plasmids, plasmids everywhere To find out more about gene edited hiPS cell lines generated...Catalog on the Allen Cell Explorer, where you can also find the certificate of analysis provided with each cell... epitope tags (e.g. FLAG and STREP) and auxin-inducible degrons (AID). Further, the Yamamoto PITCh system...on microhomology-mediated end-joining (MMEJ). To find out more about gene edited hiPS cell lines generated...

...species can be indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid...reagent/antibody, unspecified, or other. If other, indicate type in “Vector Type Other” column. For example...cloning, gene synthesis, Gibson cloning, ligation-independent cloning, restriction enzyme, TOPO cloning, unknown...or room temperature. Growth Strain Please indicate which growth strain Addgene should propagate your...in any of these standard strains, choose other, indicate strain in the “Growth Strain Other” field, and...2024.    Additional Resources on the Addgene Blog Find Articles Citing Addgene Plasmids Searchable and ...CRISPR Experiment Suggest A Plasmid You'd Like to Find at Addgene Resources on Addgene.org Start Your...

...with a common dilemma. Request the vectors you can find and use them as-is, saving time and effort but risking...available for request on the Addgene website, both as individual plasmids and as the Fragmid kit. You can also...timeline of the Golden Gate (GG) cloning approach. Individual modules and a sample ligation are depicted. (... sequencing is just part of doing good science!” Find the Fragmid kit here!  Cloning specifics Fragmid...Pol 2 protomer, Cas protein, ect...). Whatever individual components you choose will assemble in the expected...lab tested the Fragmid system by sequencing 60 individual assemblies created over five months. When they...one plasmid was the correct sequence for every individual assembly. What destination vectors are available...

... Resources on Addgene.org Find mammalian plasmids for your research Find viral vectors for infecting... hood contamination free, but you should keep in mind that UV light only kills microorganisms that it ...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...Anything between orange and light orange usually indicates that the cells have grown enough, are making their...interested in Glycobiology and Immunology. You can find Sana on LinkedIn.   References 1. Yin, R., et al...Gene expression profiling analysis of bisphenol A-induced perturbation in biological processes in ER-negative...Pagliara, Patrizia, et al. "Ostreopsis cf. ovata induces cytoskeletal disorganization, apoptosis, and gene...

...through the use of constitutive and inducible promoters. Inducible promoters are extremely useful if a...including their advantages and caveats to keep in mind before choosing a system. Mammalian expression systems...bacterial cell. Transformed cells propagate, are induced to produce your protein of interest, and then lysed..., under the control of the T7 lac promoter and induced by lactose, provides the highest level of protein...recombinant proteins under the control of the galactose inducible promoter (GAL). Other commonly used promoters ...promoters include the phosphate and copper inducible PHO5 and CUP1 promoters respectively. Yeast cells are grown...exudates Resources on the Addgene Blog Learn about inducible promoters Learn about repressible promoters Check...

...for its chance in the spotlight — Cas3. Cas3 can induce genomic alterations on a scale that other Cas effectors...capabilities of Cas3 and its partner in crime, Cascade. Find Cascade-Cas3 plasmids in Addgene's collection! A...Regardless of the subtype, the mechanism of Cascade binding and subsequent DNA degradation appears to be conserved...ensures that there isn’t a template strand left behind that the cell can use to repair. Figure ... DNA degradation continued. However, Cas3 cannot bind DNA on its own and needs to remain anchored to Cascade...throughout the process. The reeling of ssDNA and unwinding of dsDNA occur simultaneously, which creates a...reeling by the Cascade-Cas3 complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394.e3. https...

...sample for further downstream applications. As individual libraries can vary significantly, it’s important... an amplification protocol developed for each individual pooled library; generalized amplification protocols... you need 30 to 200 times as many colonies as individual plasmids in your library. As an example, if you...  Pro tip! Since you probably do not want to individually count 500,000 colonies, you’ll likely rely on... DNA, you will likely need to perform several individual transformations, such as splitting 400 ng of ...of DNA in 100 μL of cells over four individual electroporations. The specifics of this can be based on ...limiting your growth time (e.g., 16-18 hours). Finding reference protocols There are many decisions to...

...Libraries Find More CRISPR Resources at Addgene: Find CRISPR Plasmids Read our CRISPR Guide Find tools to...additional experiments investigating the role of individual genes or entire pathways in a particular disease...specific gene within the genome (see Fig. 2). Individual gRNAs are designed in silico using publicly available...Addgene, with more being added all the time. Keep in mind that libraries are only as good as the experiments...genes that confer drug resistance when mutated. Findings from this type of experiment can shed light on...phenotype. Each hit from the screen should be independently validated to ensure that the desired modification...

...software could also be used as a simple tool to rapidly find one or a few gRNAs suggested to be effective by ...knocked out at the population level, but each individual cell only has a maximum of one gene knocked out... an approach, you lose the unbiased potential to find things you are not looking for, but there are also...important for their function. Their migration is induced by various types of interactions with their environment...approved drugs already exist. If you are lucky and find that a druggable gene has a central role in your...eleven published libraries are embedded. These were kindly donated by six different research constellations...contain multiple gRNAs targeting all genes in the indicated species and were suggested to be the best gRNAs...