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We narrowed to 320 results for: POR C

Showing: 151 - 160 of 320 results

  1. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...from an additional 5–10 minutes of incubation at 37 °C. Wash the membrane for 5–10 minutes 2–3x in fresh ...stripping solution and agitate for 30–45 minutes at 50 °C. Wash the membrane for 5–10 minutes 2–6x in fresh ...directly to buffer without a re-wetting step. Important: do not dry your membrane with antibodies on it...the ones shared by Bio-Techne, ThermoFisher, or Millipore Sigma. We’ll share the basic composition of these...Alternatively, companies including ThermoFisher, Millipore Sigma, and Azure Biosystems offer pre-made stripping...your western blot several times — some researchers report up to ten cycles! Just remember that a bit of sample...

  2. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    Published
    June 2, 2020, 1:15 p.m.
    ...blog post for more details. Figure 2: Dense (c), intermediate (d), and sparse (e) labeling of the...remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed... it is an attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics... target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay, and are...Büttner’s lab designed a calcineurin transcriptional reporter consisting of a quickly-degraded destabilized ...panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate ...

  3. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ... its preferred target integration site: TNA(C/G)TTATAA(G/C)TNA. Tol2 can deliver 10-11 kB to mammalian...   References and Resources References Feschotte C, Pritham EJ (2007) DNA Transposons and the Evolution...piggyBac plasmids    Tol2  Tol2 was the first reported active DNA transposon in vertebrates. It was discovered...mutations. In these screens, the transposons encode reporter genes, mutagenic cassettes, or barcodes. When ...2021). Target site preference and host range are important to consider when selecting a transposon for a ...pronuclei of fertilized eggs, which leads to random incorporation of that sequence into the zygote’s genome, a...unpredictable. Transposons, however, efficiently incorporate into the zygote’s genome after injection into...

  4. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...application of blue light, Vvd dimerizes bringing the C-terminal and N-terminal Cre fragments together and...the PAM sequence is 5’-TBN-3’ (where B is G, T, or C). The authors demonstrate that CRISPR-Casɸ is active...in a protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed ...interest. However, FACS is unable to spatially and temporally target single cells. Now, Francois St-Pierre’...way to isolate individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures...introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim’s lab engineered...

  5. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...These plasmids enable lentiviral expression of N- or C-terminally FKBP12F36V-tagged fusion chimeras or locus-specific... rapid means of comparing functionality of N- and C-terminal fusions and degradation in a cell line of...transcription factors with important roles in Multiple Myeloma), it is important to ensure lack of degradation... chimeras For each protein-of-interest, it is important to determine which terminus can accommodate the...

  6. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...easy to switch out reporters, Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and selection...Griffith, A. L., Szegletes, Z. M., Wen, B., Kraus, C., Miller, N. W., Steger, R. J., Escude Velasco, B....New to the bench? Fragmid depositor John Doench reports even the greenest members of his lab, high school...vectors are available? What you’re building is important — but so is where you’re building it! Table 1 ...

  7. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...doi.org/10.1128/AEM.67.8.3440-3444.2001 Fogarty, E. C., Schechter, M. S., Lolans, K., Sheahan, M. L., Veseli....102684 Shareck, J., Choi, Y., Lee, B., & Miguez, C. B. (2004). Cloning Vectors Based on Cryptic Plasmids...plasmids — more on that in a minute! Fun fact! Electroporation is one popular method scientists use to promote...

  8. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ... was created by adding a 9-residue peptide to the C-terminus of human codon-optimized  cryptochrome 2 ...a fragment of the kainate receptor subunit to the C-terminus of the protein. Unlike the diffuse neuronal...while also capturing information about spatial and temporal organization. The design use to detect PKA activity...

  9. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...mutations in the common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development...1172-1179. PubMed PMID: 28671695. 6. Münch, Robert C., et al. "Off-target-free gene delivery by affinity-purified...basic research and other ongoing clinical trials support the utility of AAV as a broadly applicable gene...

  10. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...Most FP fusions for FRET studies are at the N- or C-terminus of your target protein, but even these modifications...can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition...for observed FRET efficiencies between 80% and 10%. C Cy3NTA and Cy5NTA bind His tags similarly but with...nonspecifically soak up your fluorophore package. One important concept in FRET experimental design is that the...donor first where labeling specificity is most important. Donor labeling and the need for specificity Donor...protein molecules contain the donor and, more importantly there is no fluorescence background resulting...in the ribosome, resulting in site-specific incorporation of the attached fluorophore. However, since ...
Showing: 151 - 160 of 320 results