We narrowed to 490 results for: cel.2

...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...Cellular communication and function depend on potential energy. Cellular activities like action potentials...Indicators As previously mentioned, cellular voltage is measured at the cell membrane, and this voltage is ...endocrine cells. It follows that the specific targeting of these voltage indicators needs to be cell type ...electrical events! Voltage indicators are voltmeters for cells! In this blog we will review how these indicators...related to the electrical impulse of the cardiac cells. To study this phenomenon you will likely want to...

...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...measure, and control molecular signals, cells, and groups of cells in order to understand their activity...Fluorescence-activated cell sorting (FACS): This is a type of flow cytometry that separates mixtures of cells into distinct...Thus, FACS can be used to separate cells expressing GFP from cells that are not. Developmental/transgenic... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...

...membrane integrity (Fig. 2). For example, saponin interacts with cholesterol in cell membranes and leaves ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...holes in the cell membranes of your sample to allow antibodies better access to intracellular targets. If...allowing antibodies into the cell where they could bind to intracellular epitopes. C) Permeabilizing with...method for visualizing molecules of interest within a cell or tissue. In that introduction, we broke down the...structure of your samples, the relationships between cellular components, and the accessibility of your target...cross-linkers work by (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction...

... PMID: 27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo...engineering methods have their own pros and cons (2). The NgAgo DNA guide is inexpensive and can be 5’...degraded by the cellular machinery, leaving a small amount of active NgAgo/ssDNA complex in the cell. As proposed...results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL NgAgo ARTICLE... guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA ...dangerous, non-physiological temperature for mammalian cells and one which diminishes the endonuclease activity...guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid...

...amilCP, tsPurple, and gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on... them into your cells, you can simply look for blue colonies to determine which cells have taken up the... that the cellular fitness cost varied greatly between chromoproteins. In some cases, cells containing... mScarlet as the donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you...cloning experiment worked just by looking at the cells on the plate. For example, if your plasmid contains...Here, the chromoprotein is expressed and gives the cell a magenta (amilCP) or purple (tsPurple) color. Though...added which induces expression of the toxin. Any cells that grow after this point will be those that have...

...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate...Genome-wide Inhibition Library. DNA and cell amounts After deciding on a cell type, you will need to determine...DNA and how many cells to use for your transformation(s). The amount of competent cells you need total ... generally electroporated into electrocompetent cells to ensure the highest possible transformation efficiency...efficiency. Using Stbl4 or other ultra-high efficiency cells ensures that plasmid diversity in the library is...transformation may vary depending on the particular type of cells you are using, as well as other factors, such as...transformations, such as splitting 400 ng of DNA in 100 μL of cells over four individual electroporations. The specifics...

...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...Control of Gene Expression. Cell 152:1173–1183 . https://doi.org/10.1016/j.cell.2013.02.022 Sajwan S, Mannervik...and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/10.1016/j.cell.2014.09.039 Zhou H, Liu J,...from when looking to activate genes across diverse cell lines.   dCas9-VP64 Description CRISPR activation...large and are not expressed consistently throughout cells. VPR Description VPR fuses a tripartite complex ...activation levels than SAM in some loci across Drosophila cell lines (Sajwan et al., 2019). SPH SPH combines components...

...Joachim ended up at the University of Amsterdam Chapter 2 - Joachim’s Academic and career goals. Career advice...Fluorescent Protein Plasmids 101: Visualizing Subcellular Structures and Organelles Resources at Addgene.org...Plasmids Find Plasmids for FRET Find Plasmids for Subcellular Localization ...

...days before the deadline isn’t going to be much use. 2. Originality is essential for grant success The two... it’s important not to throw in the towel if 1 or 2 funding bodies say no. If possible, get feedback from...research (either because of expertise, equipment, or excellence of the institution). Likewise, if you’re staying...Cambridge University where he studied mechanisms of cell division. Recently Seán got the entrepreneurial ...

... anniversary! We've done so much in the past two (2!) decades that it's hard to summarize, but we wanted...into the Addgene website search bar?   We'll be celebrating all year long, so keep an eye on our blog and...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing... for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify...crispr.mit.edu. They tested their system in HEK293 cells, integrating the GFP-Puro cassette into the FBL ...

...required components are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation...gone through an additional round of evolution (AID-2) to reduce some of the leakiness associated with the...uses a small molecule, a tag, and the endogenous cellular degradation machinery to function. The dTAG is...tag and small molecule. It does work in mammalian cells, mice, and even yeast, though (we have a kit!). ...and complete degradation of proteins in mammalian cells. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350...Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...

...Vanderbilt University. She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...of high profile people denying science, from celebrities falsely claiming vaccines cause autism to politicians...worried the march would be a simple, unquestioning celebration of science, at a time when scientists must rise...BC (Canada). She works in a lab that uses yeast cells to study how DNA damage promotes cancer. She is ...

...26000849. 2. Grimm, Jonathan B., et al. "A general method to improve fluorophores for live-cell and single-molecule...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...). These cell-permeable Janelia Fluor (JF) dyes have excellent properties inside living cells, especially... fluorescent proteins. In-cell labeling strategies The majority of in-cell labeling strategies have two...and oligonucleotide labels for fixed-cell imaging, not for live-cell applications. Thus, these fluorophores..."Imaging live-cell dynamics and structure at the single-molecule level." Molecular cell 58.4 (2015): 644...sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer beholden to chemists and...

...suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells. The... continuous cellular processes (e.g., protein synthesis), while amp only affects the cell wall synthesis...Check icon" %} Spectinomycin Excellent No Just bacteria Not SHuffle cells {% icon icon_set="fontawesome...icon" %} Zeocin Excellent No Bacteria, plants, and eukaryotes Not Tn5-containing cells {% icon icon_set..., penicillin, and is a beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces...

...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at...either in whole cells or as a pool of fusion proteins localized to a specific region of the cell. For example...the relationship between the age of a cell, protein, or cellular structure and a particular biological...trafficking to a subcellular location, start of gene expression, development of a cell structure, etc)....timers, researchers can look at a single image of a cell and understand how protein localization changes ...

...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....and yeast cells are cost-effective and faster-growing than mammalian cells, but mammalian cells can perform...your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture...production system, cell lines typically used include Human Embryonic Kidney (HEK) cells and Chinese Hamster...recombinant antibodies with some molecular biology and cell culture experience. Let’s go over the basics of ... place in bacterial, yeast, plant, or mammalian cells. Each of these systems has their advantages and ...

...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression...fluorescence activated cell sorting (FACS) to isolate differentiated from undifferentiated cells at the end of ...these cell types are used. The lack of good flow cytometric antibodies for many CNS-resident cell types...that drive neuronal fate and reprogramming." Cell stem cell 23.5 (2018): 758-771. PubMed PMID: 30318302...CRISPRa screening approach in embryonic stem (ES) cells to determine which transcription factors promote...antibodies, downstream of the TUJ1 coding sequence. ES cells that stably express the dCas9-Suntag activator were...

...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...entire proteome of a particular cell type, or a subset based on a cellular compartment or growth stage)....extensively to screen for molecular interactions in the cell, including protein-protein, protein-DNA and protein-RNA...field of eukaryotic transcriptional activation and cell biology. During this period, proteins were successfully...The reporter gene then allows detection of those cells where binding between the proteins of interest has...alternative fungi (C. albicans, pC2HB) (7) and mammalian cells. If the problem is that your proteins are not post-translationally...

...maybe label) your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ... for EM is not something you find in the average cell and molecular biology lab, and EM is limited in ...ultrastructural relationships that influence everything from cell division to memory, but don’t have access to the...properties of the sample itself. For example, cultured cells are typically easier to expand than mouse brain ..., Mohar B, Eddison M, et al (2022) Visualizing cellular and tissue ultrastructure using Ten-fold Robust... Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures using expansion microscopy (U-...