We narrowed to 16 results for: APP
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TypeGuide...single assay. Overview of Antibody Applications Antibody-based applications can be generally classed into ...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...of signaling molecules. Select one appropriate for your application and experimental design. Will you ...transcriptomics or proteomics. This approach is often appropriate for quantitative methods. Capture MS...protein of interest, with no overlapping epitopes. It is not appropriate for validation of an antibody...production and storage techniques, and explain common applications. Science... Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods...
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CRISPR Guide
TypeGuide... (2015). A scalable Genome-Editing-Based approach for mapping multiprotein complexes in human cells. Cell...technologies. Before CRISPR, genome engineering approaches like zinc finger nucleases (ZFNs) or transcription-activator-like...rapidly became the most popular genome engineering approach. Figure 1: Overview of the basic CRISPR mechanism... to various Cas enzymes have extended CRISPR applications to increasingly complex functions, including...understanding of CRISPR biology, introduce the various applications of CRISPR, and help you get started using CRISPR... Cas12a, which are available for specialized applications. Check out our list of additional Cas proteins...genomic edits carried out by Cas9. Multiplexing applications include editing multiple genes at once; using... -
Plan Your Experiment
TypeGuide...Cas9 in mammalian cells, many of these principles apply to using CRISPR in other organisms. First, consider...a good way to narrow down which reagents are appropriate for a given experiment. Make sure to check whether...may not be a perfect plasmid for your specific application, and in such cases, it may be necessary to customize...reagent to suit your needs. Genetic Manipulation Application Cas9 gRNA Additional Considerations Knockout ...Cas enzymes increase specificity. Dual-nickase approach increases specificity but is less efficient. Each...viral delivery of CRISPR reagents may be more appropriate. In cases where off-target editing is a major...method Expression System Components of System Application Mammalian expression vector Cas enzyme promoter... -
Guide to Using Pooled Libraries
TypeGuide...library. Apply pooled library to cells. DIVERGE: Positive and negative screens POSITIVE screen: Apply library...control sample (no selection) In parallel, apply library Apply selection; most cells will survive Perform..., semi-random sequences that can be used for applications like lineage tracing or parsing the effects ...library Apply selection Most cells die or do not pass selection (in the case of a reporter) Sort “winning...genes involved in the selection NEGATIVE screen: Apply library Perform Next Generation Sequencing (NGS)...and control cells Generate a list of gRNAs that disappear with addition of the selection mechanism Positive... -
Adeno-associated virus (AAV) Guide
TypeGuide...to be used as viral vectors for gene therapy applications. This guide contains many viral vector-specific...preferred over standard rAAV vectors for in vivo applications. Other Engineered Variants Hybrid capsids Hybrid...Vectors rAAV vectors are valuable in diverse applications due to their unique properties, including their...biomedical fields. Gene Therapy One of the major applications of rAAV vectors is in the field of gene therapy...mutagenesis. Several AAV-based gene therapies have been approved by the US Food and Drug Administration (FDA) to...neurons within the brain. Large-scale examples of mapping the brain through neuronal tracing with rAAV vectors...transgenes, enabling the development of further AAV applications. Read our blog post on split AAVs for more information... -
Cloning
TypeGuide...move around pieces of DNA. Oftentimes several approaches will work for any specific cloning project; however...likely that for any given project there is an ideal approach. This may be due to speed, cost, availability ... of time required for cloning. To utilize this approach, the fragment of DNA that you would like to clone...fragment (PCR product or synthesized oligo) with appropriate overhangs can be efficiently ligated into any... enzyme cloning. Scientists have utilized this approach to create compatible custom overhangs that can...fragments of dsDNA that have 30 or more bases of overlapping homology. One major advantage is that much larger... -
Gamma-Retroviral Vector Guide
TypeGuide... popular choice in gene therapy applications. There are many approved gamma-retroviral gene therapies ...retroviral vectors are commonly used in gene therapy applications to more specifically direct delivery of gene...lentiviral vectors, but a similar method can be applied when using gamma-retroviral vectors. Gene Therapy...cell (CAR-T) therapy is a particularly popular application of gamma-retroviral vectors. Gamma-retroviral...Gammaretroviral vectors: Biology, technology and application. Viruses , 3 (6), 677–713. https://doi.org/10.3390... -
Lentiviral Vector Guide
TypeGuide...random and difficult to predict. In therapeutic applications, like chimeric antigen receptor T cell (CAR-...Pseudotyped lentiviral vectors are common in clinical applications, like gene therapy, as you can more specifically...accurate and multiplexable method for genome-wide mapping of lentiviral integration sites. Molecular Therapy... in chromatin architecture through large-scale mapping of lentiviral integration sites. Nature Protocols...2020). An analytical pipeline for identifying and mapping the integration sites of HIV and other retroviruses...begins, by definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus... -
Chemogenetics Guide
TypeGuide...neurons) that can be turned on or off by the application of a small molecule ligand. The ideal chemogenetic...translational studies because it is FDA and EMA approved. One pitfall of a suite of muscarinic-based DREADDs... develop a PSAM that responded to a clinically approved drug, the PSAM domain was engineered to respond...Azua IR, Li JH, et al. (2009). A chemical-genetic approach to study G protein regulation of β cell function...chemogenetics for research and potential clinical applications. Science. 364(6436):eaav5282. PMID: 30872534... -
Optogenetics Guide
TypeGuide...the light source. Optogenetics has been broadly applied to study the physiology of the brain and nervous... peak expression of the opsin is reached. This approach is especially powerful when combined with site...conformation/dimerization. In addition to the applications described above, both LOV and Dronpa can control...KR, Deisseroth K. 2010 Molecular and cellular approaches for diversifying and extending optogenetics. ..., Yizhar O, Deisseroth K. 2011. Principles for applying optogenetic tools derived from direct comparative... -
Molecular Biology Reference
TypeGuide...species and can be useful for a diverse set of applications - Plasmids can drive gene expression in a wide...a primer. A primer is a small piece of DNA, approximately 18-22 nucleotides, that binds to complementary...each fluorescent signal. Sanger can sequence approximately 500-1000 bases downstream of the known primer...pieces in only a few reactions.There are multiple approaches to acquire NGS but one of the most commonly used... -
Addgene's eBook Collection
TypeGuide...contains an introduction to each vector and its applications, tips for using vectors in your experiments,...your transferable skills, and tips on the job application process. Close About Overlay Select CRISPR 101...introductions and helpful tips for antibody-based applications, from flow to IHC! Close About Overlay 0 of ... -
Adenovirus Guide
TypeGuide..., which has limited their use in therapeutic applications. However, this has also made them the most widely...Adenovirus-mediated gene delivery: potential applications for gene and cell-based therapies in the new... -
Educational Resources
TypeGuide...written and video protocols for a wide range of applications in molecular biology, plasmid cloning, and virus... -
Science Guides
TypeGuide...specific binding to other proteins. Antibody-based applications, which often use antibodies conjugated to a ... -
Promoters
TypeGuide...regulatory elements. The proximal promoter is found approximately 250 base pairs upstream from the TSS and it ...