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We narrowed to 317 results for: POR C

Showing: 201 - 220 of 317 results

  1. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...fluorophores with nonoverlapping emission spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, ... of transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the established model for reprogramming somatic...coselection, analogous to methods previously used in C. elegans, targets the sodium/potassium pump ATP1A1...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating...Genetically modified organisms like transgenic mice are important tools for understanding the function of cell types...enhancer elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e. DREADDs). The ...

  2. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ..., G. S. Parker, S. J. Qualls, B. M. Lewis, E. Ke, C. T. Dargitz, R. Wright, A. Khanna, F. H. Gage, and... Mikkers, J. Kool, L. van der Weyden, A. H. Lund, C. H. Wilson, R. Rance, J. Jonkers, M. van Lohuizen,...useful for genetic engineering approaches. Most importantly for this technology, however, is that the transposase...should be tested in the cell line of interest. Importantly, by varying these ratios, you can either maximize...obstacle, a positive selection cassette can be incorporated, and used during the differentiation process...homologous recombination, which results in the incorporation of your specific edit into the host genome. ...transposon sequences.  The use of PBx is particularly important to avoid any possible random re-integration from...

  3. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...little to no signal in the liver sample, which you do (C), so your antibody is validated! Genetic Strategies...10.1080/19420862.2015.1100787  Taussig MJ, Fonseca C, Trimmer JS (2018) Antibody validation: A view from...Reproducibility First, let’s review why validation is so important. Antibodies are a critical component of the researcher...or plan your own validation experiments, it’s important to know what you are aiming to achieve. In general...as whole cell lysate or tissue sections. It is important to confirm that your antibody is not binding to...proteins bound to the target protein, so it’s important to understand if protein:protein interactions ...

  4. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    Published
    July 21, 2015, 2:30 p.m.
    ...Plans Aren't Just More Paperwork. Vincent BJ, Scholes C, Staller MV, Wunderlich Z, Estrada J, Park J, Bragdon...and PI, it’s easy to miss out on beneficial opportunities or to find oneself unsure of the next steps....planning meetings with each trainee to create a supportive, growth-oriented lab culture. The aim of these...the overall tone of the meeting is almost more important. As DePace says, science is full of “terrible ...postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine how to...

  5. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    Published
    Oct. 31, 2019, 1:06 p.m.
    ...PubMed Central PMCID: PMC1559948. Brusca, Richard C., and Matthew R. Gilligan. "Tongue replacement in ... please! At Addgene, we love finding ways to incorporate science themes into our fun activities and Halloween...body. The fruiting bodies then rupture and release spores onto the forest below for the next unsuspecting...

  6. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    Published
    Feb. 18, 2015, 3:09 p.m.
    ...Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. An erythroid enhancer of BCL11A subject to...work was with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began...correlative biochemical characteristics and ectopic reporter assays. We have been interested in exploring the...transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via FACS, primer...

  7. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    Published
    June 26, 2017, 1:31 p.m.
    ...Addgene.   References Ke, Q., Ahn, Y., & Sugimoto, C. R. (2017). A systematic identification and analysis...physical scientists are underrepresented. It’s important to note that these results could be biased for...domains more often than other scientists   Figure 5: Portion of Tweets with Scientific Domains, Categorized ...

  8. The Michael Davidson Collection: One-Stop Shop for Fluorescent Proteins

    Type
    Blog Post
    Published
    July 29, 2014, 6:16 p.m.
    ...attach the gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest ...reagents. "We are proud to be able to bring such an important collection to the scientific community," says ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

  9. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While the unique...capsid, “pumps” the genome into the capsid through a pore in the capsid. Rep proteins are also involved in...2020) Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction. Human ...

  10. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    Published
    June 11, 2020, 1:34 p.m.
    ...annurev-pharmtox-010814-124803 Vlasov, K., Van Dort, C. J., & Solt, K. (2018). Optogenetics and Chemogenetics...   Timing Optogenetics offers a high degree of temporal resolution as receptors can be turned on and off... meet these requirements. TL;DR If precise spatiotemporal control of a small subset of neurons is necessary...

  11. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...transfection. This supernatant can be stored at 4 °C or can be directly added to your target cells. However...and plasmids.  References 1. Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging...or electrical (electric current forming membrane pores) transfection. These techniques are still used and...has been removed, Rev (which facilitates nuclear export) is expressed from a separate plasmid, and the ...you clone your gene of interest into and is an important feature for your experiment; it must be chosen...using them.   Regarding biosafety, it is also important to be aware that lentiviral particles are a powerful...

  12. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening strategy for... 22963480. PubMed Central. 5. Tsiatis, Athanasios C., et al. "Comparison of Sanger sequencing, pyrosequencing... the University of Cambridge. One of the most important steps in the CRISPR experimental process is validating...silico prediction. These techniques differ in important ways, but can also complement one another by providing... unbiased methods, high background BLESS Not reported Cell-based (in vitro) Can be used on tissue from...senstive to time since cell fixing HTGTS Not reported Cell-based Identifies translocations Limited by...

  13. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...neuronal phenotype in a mixed population (Burger, C. 2004, Aschauer, DF. 2013, O’Carroll, SJ. 2021). This...https://doi.org/10.1371/journal.pone.0076310 Burger C, Gorbatyuk OS, Velardo MJ, Peden CS, Williams P, Zolotukhin...fibrillary acidic protein (GFAP) for astrocytes it is important to test it in the brain region of interest, especially...of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse... the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When Using AAVs Three Key Considerations...

  14. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    .... DOI: https://doi.org/10.1080/rsob.180241 Vanni, C., et al., Unifying the known and unknown microbial...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as... screens to introduce reporters throughout an organism’s genome. These reporters have historically been...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...promoter – all rely on the random integration of a reporter construct in close proximity to an endogenous ...example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed in the kidneys... for with an uncharacterized gene. The actual reporter construct can be something phenotypic (such as...

  15. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...of our most popular mammalian expression vectors, c-Flag pcDNA3, comes from the pcDNA series. It has the...consider as you make your choice. The first and most important thing you need to know is your expression system...have to make sure that your plasmid has been incorporated into the host organism, usually achieved with...basal expression level, induce expression at an opportune time or go all out with constitutive high level...Some of the more popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. ...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...

  16. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ... Deverman BE, Matho KS, Cetin A, Woodard K, Cepko C, Guerin KI, Rego MA, Ersing I, Bachle SM, Kamens J...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...the Cre recombinase, are used to spatially and temporally control transgene expression. Lox sites are directional...sites lets researchers control the spatial and temporal activation of a transgene. Figure 2: FLEX...structure when transcribed which aids in nuclear export of mRNA and helps increase the expression of your...polyadenylation or polyA signal aids in the nuclear export of RNA and RNA translation, and promotes RNA transcript...

  17. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    Published
    March 28, 2022, 4:30 p.m.
    ...Boeckman JX, Manning CF, Nguyen JT, Bechtold H, Dumitras C, Gong B, Nguyen K, van der List D, Murray KD, Engebrecht...and have always been, a community resource that supports both toolmakers and tool users. We encourage you...Educational Resources As part of our commitment to supporting your research, Addgene is creating an array of...Through our incredible community, including your support, we have enabled scientists to easily share plasmids...Studies Hybridoma Bank, a nonprofit that has been supporting hybridoma sharing within the community for decades...Immunofluorescence Addgene’s Antibody Guide The project is supported by a NIH BRAIN Initiative grant (U24NS119916 )...

  18. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    Published
    Jan. 22, 2019, 2:41 p.m.
    ...∆G, however, can not be conjugated to PE due to a C-terminal glycine deletion, so it remains in the cytosol... development of several fluorescent autophagy reporters now allows for the measurement of autophagic flux...makes Keima useful for monitoring mitophagy. It’s important to remember that because Keima’s fluorescent relies... al. "Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...using a single fluorescent biosensor." Scientific reports 5 (2015): 12397. PubMed PMID: 26215030. PubMed ...

  19. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ... antibodies aren’t free to bind endogenous IgGs. (C) A third solution is to use Fab fragments against ...primary antibody that is directly conjugated to your reporter circumvents the issue. However, directly conjugated...which uses just a primary antibody that has your reporter (in this case a fluorophore) directly attached... as effective for other techniques, but it is important to be aware that the issue is not unique to IHC...

  20. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    Published
    Feb. 13, 2018, 2:51 p.m.
    ...released from nuclei by a short incubation at 37 °C. These short DNA fragments can then be purified for...MNase to antibody bound proteins (Figure 1). Importantly, since Protein A has weaker affinity for mouse... rather than to rely on nuclei extraction. They report that this change offers a significant improvement...
Showing: 201 - 220 of 317 results