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We narrowed to 295 results for: SAM activators

Showing: 241 - 260 of 295 results

  1. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ...homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson...by intracellular factors such as nucleases and reactive oxygen species, or external forces such as ionizing... occur; however, if both HJs are resolved in the same manner, this results in a non-crossover event.  ...

  2. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...effect within the body, or they can be used to activate cellular pathways by binding to cell surface receptors... exact amount of protein in your sample, you’ll compare your samples to a standard curve. This curve is...the known samples is used to calculate the concentration of protein in the experimental samples. Get more...multiple proteins on the same cell simultaneously as well as compare between samples. Learn more about flow...of different isotypes can be used together in the same experiment because the reagents used to detect the... population of antibodies that all recognize the same epitope of the target protein. Most commonly these...expression in cell lysates, whole cells, or tissue samples. Many experiments that use antibodies use a primary...

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

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...expressing TH protein indicating that there is promoter activity. Additionally, some neurons express low levels... with the same titer and serotype previously used in a different brain region that the same result will...sites will be flanked by two lox sites now in the same orientation. Next, the loxp site is excised since...since it is between two lox2272 sites in the same orientation preventing the transgene from reverting. Figure...scrambled one pair of lox sites and either used the same sequence to flank a transgene (homologous pairs)...the DIO/FLEx vector. This step is essentially the same as what was used to optimize the serotype, except...

  4. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ...chemogenetic tools, where modification of cellular activity outside of your experiment site or circuit can... different phenotypes of the same cell type, are intermingled in the same brain nucleus: a serotype that...tropism between animal species or even strains of the same species. What works well in a mouse may not work...doi.org/10.3390/cells12050785 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term Doxycycline-Regulated...10.3390/pharmaceutics13050750 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...

  5. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ... hybrid vector, as measured by β-galactosidase activity, was 81% of that from a single vector, but significantly...nbt0702-697 Hirsch ML, Agbandje-McKenna M, Jude Samulski R (2010) Little Vector, Big Gene Transduction:...Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV Transductifon Is Mediated...

  6. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...far-red genetically encoded indicators of neuronal activity. J Neurosci Methods 362, 109314 (2021). Yu, D...., and deep penetration of red-shifted light. The same NIR FPs and biosensors can be imaged at spatial ...

  7. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...RNA cleavage, and translational repression or activation: Kellermann, Stefanie J., and Andrea Rentmeister...plasmids used to generate these fusions all use the same restriction enzymes for cloning. Furthermore, some...

  8. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    Published
    Nov. 6, 2015, 3:30 p.m.
    ... metastasis as well as teaching and mentoring activities. Follow her on twitter @zcastan or on linkedln... my mentor, since she does research at UK in the same field that I do: prostate cancer. She has guided... Pilot Program 2015-2015 Scholars need strong proactive mentors who have relevant expertise and skills...

  9. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...Griesbeck, Heim, & Tsien, 1999), monitoring kinase activity (Ni, Titov, & Zhang, 2006), quantifying antibody... However, the acceptor doesn’t need to go to the same exact location as the donor. As long as some of ...targeted nearby (such as a neighboring house on the same street as the donor), then FRET measurements are...Ebright, 2001). These Cy/NTA conjugates work the same way as NTA-sepharose beads used for protein purification... binding sites are adjacent to each other on the same protein and a negative control experiment conducted...

  10. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...effects, each group finds very low to no off-target activity at the ten highest-ranking predicted off-target...slightly different methods, they each observed the same positive phenotypes in cardiac and skeletal muscle...

  11. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ... design has been shown to increase T7 promoter activity. To avoid re-cutting by Cas9 after editing, make...They have successfully used guides targeting the same or complementary strands, as well as guides facing...

  12. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...protein level at the timepoint studied Off-target activities of the RNAi Correctly interpreting your data...the abundance of the epitope in the sample, relative to cross-reactive antigens – that is, an antibody shown... recognize different epitopes of the same protein, show the same pattern of labeling, this is supportive...detecting the target-of-interest in your assay and sample-of-interest. The approaches available broadly map...determining the selectivity of antibodies for other sample types and for non-surface (intracellular) antigens...approaches used will be different depending on the sample and target type. To start the validation process...the target at a robust level, similar to that in samples of interest. If you are working with cells that...

  13. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ...to researchers.  TALENs In 2010, transcription activator-like effector nucleases (TALENs) were engineered...findings of the 1952 Hershey-Chase experiment. In the same year, Rosalind Franklin and her student Raymond ...

  14. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...Resources on the Addgene Blog Learn to Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate...investigator can be relatively certain that their samples don’t have off-target mutations if they remain ...

  15. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...spatiotemporal control of transgene expression or activity can be achieved using switches, including riboswitches...function like your average AAVs — and have many of the same restrictions, like limited cargo space (~4.7 kb)...systems, which use two or more AAVs to transduce the same cell, can accommodate larger, modular genetic payloads...efficient AAV-PHP.eB, developed and deposited by the same lab. AAV-PHP.eB is the most commonly requested systemic...off-target organs. AAV.CAP-B22 was developed through the same process as AAV.CAP-B10 and shows high transduction...

  16. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ... the sample to that of a standard curve to extrapolate the concentration of antigen in the sample. A variety...with samples, eliminating them from the procedure removes one potential source of cross-reactivity.   While...use the same secondary antibody for any ELISA protocol that uses primary antibodies of the same isotype...-containing sample with their conjugated primary antibody. Users next incubate the sample with the multiwell...used to quantify the level of target antigen in a sample. While Engvall et al. originally developed the ... proteins and small molecules from a variety of sample types (Engvall 1971). Scientists value ELISAs for...using the method. ELISA basics All ELISAs follow the same overall process. First users coat a multiwell plate...

  17. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    Published
    May 23, 2023, 1:15 p.m.
    ...presence of several biomarkers together in the same tissue sample predicts a certain cancer and want to look...you will need to start with a fixed sample. Depending on the sample type, this might be frozen or formalin-fixed...you image the first target, you inactivate the fluorophores in the sample, either by chemical treatment ...al, 2022 for a sample protocol.)  Simultaneous IF Depending on your needs, your sample, and targets, you...known to be abundant in your samples (with a primary antibody that has the same species and isotype as the...set of samples where you label only one target in each. Compare these individually-labeled samples with ...the sample is being damaged by all the handling steps, or there’s some unexpected cross-reactivity. Keep...

  18. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ...than generally reactive ones. And make sure to check for any reported cross-reactivity when planning your...minimize the risk of cross-reactivity. To further limit species cross-reactivity, try to find secondary antibodies...cross-reactivity. For example, if you are using mouse IgG2a and rat IgG1 antibodies in the same application...specific secondary antibody, rather than a broadly  reactive secondary (Fig. 1). For example, an anti-mouse...Manning 2012).      Figure 1: Broadly reactive versus subclass-specific antibodies. Only a fraction...of the antibody molecules present in a broadly reactive secondary antibody will bind to a monoclonal primary...different isotypes and subclasses, use a broadly reactive secondary antibody to increase the chance of primary...

  19. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ... replicates are repeated measurements of the same sample and are included to assess variability in the... the tau western blot several times with the same samples and compare the results across runs. If the ... wild-type and knockout samples. In this test, wild-type and knockout samples are treated with the identical...present in the wild-type samples and absent in the knockout.  If knockout samples are unavailable, try substituting...control, and cross-reactivity control when multiplexing. A, wild-type and knockout samples are the gold standard...interactions between the sample and the secondary antibody. C, a label control treats the sample to the full protocol...interpretation of results. A positive control is generally a sample or group that will have a desired response. For...

  20. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    Published
    July 14, 2021, 1:15 p.m.
    ...recognize the same region, or epitope, of the target protein. These antibodies are also all the same isotype...heterogeneous mixture of many antibodies that recognize the same protein. In the immune system, antibodies are produced...peptide sequence derived from this protein. At the same time, an adjuvant such as KLH or Freund’s adjuvant... without causing high background due to cross-reactivity. When to use polyclonal antibodies One advantage...clones will still recognize your protein. For the same reason, they’re also useful in experiments where...experiment, you should either order several vials of the same lot of polyclonal antibody, or select a monoclonal...your polyclonal antibody, you’ll want to use the same positive control across experiments to be aware ...
Showing: 241 - 260 of 295 results