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We narrowed to 898 results for: NSI

Showing: 661 - 680 of 898 results
  1. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... In research, iodixanol is used as an isomolar density gradient medium suitable for virus purification...to mammalian cells. Therefore, and unlike other density gradient media, it is reportedly not necessary ...steps are used to remove contaminants with lower densities, including empty capsids. The 60% step serves ...
  2. Antibody Guide

    Type
    Guide
    ...Figure 1. The two arms of the Y structure are responsible for binding the antigen, and are called the antigen...isotype of the antibody. B cells, the immune cell responsible for producing antibodies, can change the constant...binding site. In B cells, the genetic region responsible for the variable region goes through a randomized...valency of 2 IgE Protects against parasites and is responsible for driving allergic reactions such as anaphylactic...requirements, monoclonal antibodies are typically more expensive than polyclonal antibodies. Environmental factors... Minibodies are engineered antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments... protein. Fluorophores Fluorophores are light-sensitive molecules which emit a specific wavelength of ...
  3. Plan Your Experiment

    Type
    Guide
    ...Manipulation Application Cas Enzyme Additional Considerations Browse Plasmids Knockout Permanently disrupt...essentially function as a “knockout,” but be sure to consider potential consequences of having a non-functional...be no PAM site close to your desired edit site. Consider alternative Cas9 enzymes with different PAM sequences...completely avoid them. To increase specificity, consider using an engineered Cas enzyme with higher fidelity...to off-target activity, it is also important to consider factors that maximize cleavage of the desired ...with BioRender.com. When choosing your system, consider which Cas enzyme you are using (as species and...for your experiment) and the expression time (transient or stable). For plasmid and viral-based approaches...
  4. Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs

    Type
    Protocol
    ..., be aware that some gloves may contain latex. Consider nitrile gloves as an alternative. For extra precautions...anyone else associated with Addgene, can be held responsible for your use of the information contained in ...
  5. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...satellite colonies will grow). It is, however, more expensive. With a 6 cm diameter petri plate containing solidified...Plate 50 µL of transformed E. coli /rescue media suspension onto the agar and gently spread over the surface...
  6. Centrifugation

    Type
    Protocol
    ...from a liquid or separate liquids of different densities is essential to many different experimental procedures...samples. If your samples are significantly higher density than water, then you may need to fill the blank...
  7. Gibson Assembly Protocol

    Type
    Protocol
    ...and molecular biologists now use this method extensively. Why Gibson Cloning? No need for specific restriction... Assembly master mix (isothermal reaction mix) consists of three different enzymes within a single buffer...
  8. General Transfection

    Type
    Protocol
    ...linear polyethylenimine. Transfections allow for transient expression of a gene of interest in a target cell...discard the tube and thaw a new working stock. Considerations Before You Start The health of the cell line...
  9. Immunocytochemistry

    Type
    Protocol
    ...well. Allow the HeLa cells to grow to the desired density before labeling. Section 2: Fixing and permeabilizing...instructions before starting your experiment and consider titrating your antibody to determine the optimal...
  10. Optogenetics Guide

    Type
    Guide
    ...activated. In the past, the field of optogenetics was considered to include both actuators (genetically-encoded... the following enhancements: Increased light sensitivity Example: ArchT Increased photocurrent amplitude... Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum 500 Chrimson...from Halorubrum strain TP009. Improved light sensitivity over Arch. 566 eArchT 3.0 ArchT with addition... delivery system. Here are some key factors to consider: Excitation or inhibition? First things first:...neural populations at distinct times. Temporal considerations Temporal precision is key in optogenetic experiments...Rhodopseudomonas palustris bacteria. Key feature is the sensitivity to 740- to 780-nm near-infrared light. 740–780...
  11. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...the viral genetic material inside. The AAV genome is ~4.7 kb long and consists of two open reading frames...ITRs is limited by the physical space available inside of the small AAV capsid and directly correlates...Genome Integration Recombinant AAVs are generally considered to not integrate into the host genome. When wild-type...during production, as it carries the Cap region responsible for the production of capsid proteins. AAV2 is...it was the first to be well-characterized and extensively studied, and has a broad tissue tropism. Table...information. Self-complementary AAV (scAAV) One downside of the use of AAVs as viral vectors is their single-stranded...common points of entry. Biosafety should always be considered with respect to the precise nature of experiments...
  12. Chemogenetics Guide

    Type
    Guide
    ...molecule ligand. The ideal chemogenetic tools are unresponsive to native ligands and are engineered to respond... development of mutated receptors that while unresponsive to endogenous ligands, could be activated by... coupled receptors, but unlike RASSLs, show insensitivity to endogenous ligands, have low constitutive...proteins of a light-emitting luciferase and a light-sensing optogenetic element. When the luciferase substrate...luciferase-fluorescent protein fusions, to more-sensitive opsins has led to improved LMOs with increased...chemogenetics experiment, here are some things to consider: Activation or Inhibition? Do you want to turn... a study in rats using an hM4Di receptor might consider an alternative DREADD ligand such as DCZ versus...
  13. CRISPR Guide

    Type
    Guide
    ...fusing the anti-CRISPR protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled...There are two main types of base edits: Base transitions — edit to same classification of nucleotide base...pyrimidine-to-pyrimidine), which catalyze base transitions. Cytosine base editors are created by fusing ...integrated along with the sequence, so the edit is not considered “scarless”. Figure 7: Overview of CRISPR transposases... target gene function. It has also been used extensively to screen for novel genes that regulate known...simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single transcriptional...silencing across much of the human genome following transient expression of the dCas9 fusion, with the epigenetic...
  14. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... 96-well optical plate Pipette tips General Considerations Always run standards and samples in duplicate... #59462 is one plasmid that gave reliable and consistent results. Use the recommended plasmid, or test...
  15. Pouring LB Agar Plates

    Type
    Protocol
    ... bath Pipetman Reagents 37 g pre-mixed powder consisting of: 5.0 g yeast extract 10.0 g peptone from casein...colonies will grow). However, it is also more expensive. Procedure Measure 37g of pre-mixed LB-agar powder...
  16. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...After staining with Coomassie blue, protein band intensities are measured using ImageJ software and a standard...similar photo software to determine the relative intensity of the protein bands to the overall lane. Import...
  17. Western Blot

    Type
    Protocol
    ...blocking buffer and optimal antibody concentrations. Consider titrating your antibody to determine the optimal...instructions before starting your experiment and consider titrating your antibody to determine the optimal...
Showing: 661 - 680 of 898 results