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We narrowed to 587 results for: cel.2;

Showing: 331 - 340 of 587 results

  1. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...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...

  2. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...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...

  3. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ... 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...

  4. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...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...

  5. Pooled Library Amplifications

    Type
    Blog Post
    ...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...

  6. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...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...

  8. Twenty Years of Sharing Science!

    Type
    Blog Post
    ... 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...

  9. 6 Tips for Grant Writing

    Type
    Blog Post
    ...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 ...

  10. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ... 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 ...
Showing: 331 - 340 of 587 results