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We narrowed to 569 results for: CIT

Showing: 81 - 100 of 569 results

  1. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be affected by...emitted by a Renilla luciferase variant are used to excite the fused fluorescent protein. This technique is... used FRET, but it eliminates the need for an excitation light source. Instead, a luciferase substrate...-lanterns are not subject to photobleaching/phototoxicity, they successfully imaged multiple subcellular...constant supply of substrate. Using this compound to excite the Nano-lanterns, Takai et al. continuously imaged...imaging, especially where photobleaching and phototoxicity are a concern, these constructs come in very...complement optogenetic tools, where light used for excitation can cause unintended activation of the optogenetic...

  2. A Control for All Seasons

    Type
    Blog Post
    ...primary antibody specificity. The gold standard in demonstrating primary antibody specificity is to test the...include a primary antibody specificity control, secondary antibody specificity control, label control, and...Secondary antibody specificity  In indirect staining approaches, secondary antibody specificity must also be... be able to confirm that the testing protocols elicited the expected response and have a baseline with... antibody-based experiments Primary antibody specificity Antibody-based assays have their own unique controls...gold standard to demonstrate primary antibody specificity. B, secondary antibody-only staining will allow.... Non-specific binding  Primary antibody specificity is further supported by a pre-immune serum or...

  3. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...without changing excitation wavelength (Salomonnson et al. 2012). They can also be excited efficiently at...bone marrow to the thymus where they gain their specificity or from the thymus to the lymph nodes. These ...source is mounted above (epi) the specimen and the excitation light passes through the microscope objective... or dyes. 3. Two-photon microscopy Two-photon excitation microscopy is a fluorescence imaging technique...fluorescent molecule (see figure to the right). The excitation generated by these two-photons occurs only at...physio-pathological challenge. However technical specificities have to be taken into consideration especially...commonly used orange and red fluorescent proteins are excited by 750 nm to 760 nm laser light, enabling dual ...

  4. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...where labeling specificity is most important. Donor labeling and the need for specificity Donor fluorophores...fluorophores. When the first fluorophore (the donor) is excited, it can transfer energy directly via resonance ...FRET fluorophores? When is absolute labeling specificity needed and when is it not? What are the proper...proper mailbox (i.e., a binding site) in the right city (protein). In a cell, this task is even harder due...FRET experimental design is that the targeting specificity requirements are different for the donor and ...single mailbox at a specific house in your molecular city). However, the acceptor doesn’t need to go to the...are delivered to the wrong address in a different city, you can still develop a successful FRET-based assay...

  5. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...Packaging Capacity (>8kb) High Transduction Efficiency Transient Gene Expression High Immunogenecity ...the brain. Secondly, some viruses have a low immunogenicity which makes them ideal for applications with...primary advantage over adenovirus is their low immunogenicity, which means they do not stimulate a strong...attractive tool for human subject delivery, as immunogenicity was previously a large barrier for gene therapy...Tropism Only Infects Dividing Cells Moderate Immunogenicity Insertional Mutagenesis Possible Lentivirus...Episomal (Little Risk of Mutagenesis) Very Low Immunogenicity Infects Non-Dividing Cells Transient Gene...Mouse Cells and Hematopoietic Cells Low Packaging Capacity (~4.5kb From basic science to clinical applications...

  6. Bringing Renewable Energy to Addgene

    Type
    Blog Post
    ... of an “electricity aggregation program” for all local households and businesses. Electricity aggregation...September all will be powered by 100% renewable electricity! A (not so) shocking transition  In early July...program offers options of between 14% - 100% electricity supplied from renewable sources and we have chosen... As a result, we will be drawing 100% of our electricity from renewable energy projects right here in ...save energy. The transition to 100% renewable electricity will allow us to maintain these particularly ...more eco-friendly The switch to 100% renewable electricity is certainly not the only sustainability effort...

  7. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...of the page that help you cite this publication in your own work: How to cite this plasmid: The Recipient...template citation for you to use in your Materials and Methods and References sections when citing the use...Acknowledgement (RSA) asks requesting scientists to cite the depositing lab whenever their plasmids are used...Addgene catalog number to easily find template citations for Addgene plasmids you have used in your research...including genbank files and protocols here. Articles Citing this Plasmid: This is a relatively new feature ...Resources on the Addgene Blog Read about Our New Citations Feature Check out Our Walkthrough of the Deposit...

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

    Type
    Blog Post
    ...protocolLess specificityIncreased chance of cross-reactivity Sandwich High specificityHigh sensitivity... Scientists value ELISAs for their accuracy, specificity, and sensitivity (a good ELISA can detect picogram...). Fluorescent tags may also be used, but for simplicity we will discuss the more common enzyme-based ... primary antibodies of the same isotype.     Specificity and timing are the major disadvantages of the...capture and detection steps, which increases the specificity of the assay. The assay is also flexible, as ...of background stainingLess sensitivityLess specificityLimited availability of conjugated antibodies ...of cross-reactivity Less sensitivityLess specificityLimited availability of conjugated antibodies ...

  9. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...retroviruses generally have moderate to high immunogenicity, meaning they induce an immune response in ... cargo into a cell. These viruses have a cargo capacity of ~ 8 kb, ample room for most experimental needs.... Lentiviruses have slightly larger packaging capacities than gamma-retrovirus, coming in around 9 kb....along with the endogenous DNA. AAV also has low immunogenicity, meaning it does not induce the same high immune...direct application of virus. However, the cargo capacity of AAV is ~4.7 kb, almost half that of lenti and...identified so far! Adenovirus generally has high immunogenicity, but some strains have been engineered to induce...too. May your titer always be high and your immunogenicity low! References and Resources References Robinson...

  10. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...Energy Transfer. Unlike the typical excitation and emission of an excited fluorophore, FRET involves a non-radiative...Donor fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly...transfer of energy (i.e. no emitted photons) from the excited donor fluorophore to the acceptor fluorophore. ...spectrum of the donor fluorophore overlaps with the excitation spectrum of the acceptor fluorophore. Otherwise... transferred from the donor will not be able to excite the acceptor. In addition to sufficient spectral...

  11. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...monomeric cyan-excitable red fluorescent protein (mCyRFP1). Derived from cyan-excitable organge fluorescent.... While certain cell types, such as principal excitatory cells and glia, have been successfully targeted...fluorescent protein (CyOFP1), some of the exciting properties of mCyRFP1 include a redshifted-emission spectrum...major advantage of mCyRFP1 is its ability to be coexcited along with EGFP while retaining an emission spectrum...CaMKII and RhoA during induction of structural plasticity in single dendritic spines. The set of CyRFP ... David Liu’s lab increase both the range and specificity of base editing. Kim et al. used natural and ...on-target editing efficiency. To further improve specificity, they purified HF-BE3 protein for delivery in...

  12. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...with broad CNS or PNS transduction, cell type specificity, or retrograde transport in neurons. (b) Genetic...by triangles] facilitate tissue or cell type specificity. An on/off genetic switch can be used for expression...susceptible to off-target transgene expression-induced toxicity in NHPs (Hordeaux et al, 2020; Hudry et al, 2023...designs and administration doses. Note: If capsid immunogenicity is a concern, using a purification method that...broad):  enhanced coverage for inhibitory versus excitatory neurons Mice, rats Yes (LY6A) Chan et al...AAV.CAP-B10 CNS: balanced coverage of inhibitory and excitatory neurons Mice, marmosets Unknown Brown et...capsids were deposited by the Gradinaru lab. For simplicity, in each section we’ll list the Gradinaru capsids...

  13. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...showed that Cas7-11 cut RNA with unprecedented specificity: when purified, it could be programmed to generate... targeting experiments. If Cas7-11 lacked this toxicity, it would represent a major step forward for CRISPR-based...up the potential of further development of low-toxicity RNA editing tools.   Fig. 1 Schematic of...the significantly reduced amount of off-target toxicity compared to other RNA editing methods — streamlining...viability – meaning there was no measurable cell toxicity – using Cas7-11. This included tests done in cell... cell lines known to be more sensitive to toxicity from RNA editing, such as mouse embryonic stem cells...CRISPR has yet to fully taken advantage of, but exciting new tools like these represent a major step forward...

  14. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...novel SpCas9 variants with altered PAM binding specificity: SpCas9 variant Mutations (relative to ...with an NGA PAM. This variant may increase the specificity of genome modifications at DNA targets adjacent...expanded PAM recognition as well as increased specificity and lower off-target activity (Hu et al., 2018...Kim J-S (2016) Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol...Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature 523:481–485 . https://doi.org/10.1038...McCaw ZR, Aryee MJ, Joung JK (2016) Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells....variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 . https://doi.org/10.1038/nature26155...

  15. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    ...have high activity and specificity with low cross-reactivity, as well as low toxicity. FLP/FRT recombinases... ligand binding domain-fusion can prevent this toxicity, but with the caveat of partial rather than complete.... Finally, Rubin’s group assayed the potential toxicity of these recombinases by crossing flies homozygous...carried both Cre and GAL4, further emphasizing the toxicity of this enzyme in Drosophila. If you're still ....161269798. PubMed. Reduction of Cre recombinase toxicity in proliferating Drosophila cells by estrogen-...

  16. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ... to no toxicity in the knockout line, it is likely a specific drug with low off-target toxicity. This ...candidates are isolated, they can be validated for specificity to their target, or their direct target can be...developed, it’s good practice to measure the specificity of that target and validate the hit compound....toxic to both lines, it probably has either high toxicity or high off target effects, both of which make...keeps advancing, so will drug discovery – it’s an exciting time to be in the biomedical research field! ...

  17. Important Considerations When Using AAVs

    Type
    Blog Post
    ...interest. Although they have a small packaging capacity, AAV vectors were suitable for our needs for the...vivo animal model should consider the possible elicited immune response from viral infection. Animal safety... also have similar transport characteristics (velocities of movement, distance traveled, etc.) (4). This...identified with AAV2 (5), AAV6 (6) and AAV5 at excitatory connections (3). These facts beg the question...potential of AAVs is their ability to be used with new exciting technologies (such as CRISPR/Cas9) as they allow...Joshua C., and Richard J. Samulski. "Packaging capacity of adeno-associated virus serotypes: impact of...recombinant adeno-associated viral vectors: packaging capacity and the role of rep proteins in vector purity....

  18. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Editing Specificity” by Tycko et al. 2016 and “Defining and improving the genome-wide specificities of CRISPR–Cas9...have been widely adopted due to their relative simplicity and low cost. The problem with these assays is...cleavage reveals RNA-programmed Cas9 nuclease specificity." Nature biotechnology 31.9 (2013): 839-843. ...genome-wide analysis of zinc-finger nuclease specificity." Nature biotechnology 29.9 (2011): 816-823. ...Joung. "Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases." Nature Reviews Genetics17.5...Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity." Molecular Cell 63.3 (2016): 355-370. PubMed...

  19. Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

    Type
    Blog Post
    ....   Excitation with NIR light also produces less non-specific ROS because NIR light doesn’t excite natural...2I-dL5**).   A FAP-TAP complex will light up if excited with light of specific wavelength and also produce...  The iodine-modifications also shift MG-2I’s excitation wavelength to 666 nm, which is within the near-infrared... non-specific ROS when exposed to YFP and RFP excitation wavelengths, which suggests that it could be ...increase cell ablation efficiency, but dL5** phototoxocity is induced when expressed on the cell surface...

  20. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...photobleaching and phototoxicity that standard FPs might induce though their continuous excitation. By generating...chromophore which contribute to the 2 peaks of wtGFP’s excitation spectrum- a major 397 nm peak and minor 475 nm...anionic form, causing increased fluorescence when excited with 488nm blue light. The T203H mutation in PA-GFP...Contrast (Fold Change in Fluorescence) Brightness Excitation/ Emission (nm) PA-GFP Monomer UV-Violet Dark...Contrast (Fold Change in Fluorescence) Brightness Excitation/ Emission (nm) Dronpa Monomer UV-Violet Dark...own quirks (level of zoom, intensity, possible excitation lasers) it is also important to optimize your...
Showing: 81 - 100 of 569 results