Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 927 results for: TIM;

Showing: 101 - 120 of 927 results

  1. Antibody Guide

    Type
    Guide
    ...conjugate or conjugation process. Sodium azide, an antimicrobial agent, can block some amines needed for conjugation...In those cases, thimerosal can be used as an antimicrobial agent. Other chemical interferences with conjugating...measurable light or color in response to a specific stimulus, can be conjugated to antibodies in order to allow...produces a colored product that accumulates over time which is relative to the amount of enzyme present...purchased already conjugated, which can save significant time in the lab. However, if the antibody you want is...ubiquitously expressed protein such as GAPDH or tubulin (sometimes referred to as housekeeping genes). This allows...protein or protein interactions in a specific timeframe or context. Immunoprecipitation (IP) IP uses immobilized...

  2. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...10.1093/nar/gku936 Eva K. Brinkman, Arne N. Kousholt, Tim Harmsen, Christ Leemans, Tao Chen, Jos Jonkers, Bas...into your target cells with Cas9. Hooray! Now it’s time to make sure your genome edits went according to...within a given genomic window, along with the estimated editing frequency of your sgRNA-Cas9 combo. Pro... from the donor DNA. This method can precisely estimate editing frequency in a bulk population and also...Cas9 or gRNA is the issue, check out or blog on optimizing this step, or consider enriching for transfected...

  3. Optogenetics Guide

    Type
    Guide
    ...neurons upon exposure to light through direct stimulation of ion channels. Naturally occurring channelrhodopsins... nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely ...been created via genetic point mutations, codon optimization, and chimeric fusion of domains from two different...engineered with enhancements such as: Human codon optimization Example: Halo Increased photocurrent amplitude...activate different neural populations at distinct times. Temporal considerations. Temporal precision is ...and promoter system used, there is an incubation time (days to weeks) until peak expression of the opsin...fused to Cry2 and the other half to CIB1, light-stimulated heterodimerization can reconstitute the protein...

  4. CRISPR Guide

    Type
    Guide
    ... an optimized gRNA for your application (see: Plan Your Experiment ). In addition to an optimized gRNA... currently no defined measure of fidelity, so sometimes they will be referred to as increased fidelity...structured motif to enhance pegRNA stability PEmax - optimized PE architecture; increased synergy between PE ...compared to HDR. Prime editing requires more optimization and typically has lower efficiency (< 50%) than...integrase landing site ( attB ) and the other to stimulate DNA repair. This can result in more efficient .... gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high on-target activity...activators and targeting promoter regions. You might sometimes see these referred to together as CRISPRi/a. dCas9...

  5. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ... indicating the optimal serotype(s) for transduction of a given organ. Tissue Optimal Serotype Central...viral replication (E4, E2a and VA). These are sometimes combined within the packaging plasmid. For a summary...occur, although at a low frequency which has been estimated to range from 0.01% to 3% depending on cell type... range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue types, like... lab has developed plasmids containing SaCas9 optimized for their use in rAAV vectors . Read our blog ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...protein (DBP) that enhances DNA replication by stimulating elongation and stabilizing the displaced single-stranded...

  6. Molecular Biology Reference

    Type
    Guide
    ...field of recombinant DNA technology have been optimized for their use of studying and manipulating genes...bacterial cell division is reduced (i.e., it takes more time to copy this extra DNA). Because of this reduced...form of another antibiotic resistance gene (this time, under the control of a non-bacterial promoter) ...origin of replication, and MCS. They are small and optimized to help in the initial cloning of a DNA fragment...developed the process termed Sanger sequencing, sometimes referred to as chain-termination sequencing or...Sanger sequencing requires a lot of DNA because the ultimate goal is to have a fluorescently tagged nucleotide...captures the fluorescent color that is emitted each time a base is added. Again, each base (A,C,T, or G) ...

  7. Cloning

    Type
    Guide
    ... over the years to move around pieces of DNA. Oftentimes several approaches will work for any specific...specific DNA sequence, which can make it difficult at times to identify compatible restriction enzymes, that...behind a short scar in the DNA sequence and can be time consuming compared to other cloning methods. You...simplifying the process and reducing the amount of time required for cloning. To utilize this approach, ...proteins. Thus, you can clone your gene of interest one time into a donor plasmid (or acquire one that already...polymerase used, and the single A overhangs degrade over time, further reducing ligation efficiency. TOPO® ready...

  8. Plan Your Experiment

    Type
    Guide
    ...putative target sites have been identified, it is time to choose which site is likely to result in the ...Using validated gRNAs can save your lab valuable time and resources when carrying out CRISPR experiments...Once you have selected your target sequences it is time to design your gRNA oligos and clone these oligos...proceeding with your experiment, it may be necessary to optimize your delivery conditions. Learn more about CRISPR... gRNA and Cas enzyme to your target cells, it is time to validate your genome edit. CRISPR editing produces...

  10. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...start sites, such as promoters and enhancers. Many times, integration sites are found in proto-oncogenes.... of helper-free production cell lines also saves time in downstream purification, as extraneous plasmid...different retroviral vectors using a novel, CoDOn-Optimized gene for chimeric GALV envelope. Viruses , 13 ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...

  11. Adenovirus Guide

    Type
    Guide
    ...size of the DNA removed from the viral genome. Over time, subsequent generations of rAdV vectors have been...needing transgenes to express for a shorter period of time or when needing to package larger cargo. CRISPR-...vectors are also being used in cancer therapy to stimulate anti-tumor immune response in different ways, ... a given virus or infectious agent at a specific time point. Transduction The process of artificially ...

  12. Lentiviral Vector Guide

    Type
    Guide
    ...assess viral integration (see Kim et al. (2021), Timms et al. (2018), and Wells et al. (2020) for examples..., A., & Pour, M. F. (2020). Concise review on optimized methods in production and transduction of lentiviral...https://doi.org/10.1261/rna.2192803 PMID: 12649500 Timms, R. T., Tchasovnikarova, I. A., & Lehner, P. J. ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...

  13. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...27760321. PubMed Central PMCID: PMC5081405. 6. Wang, Tim, et al. "Gene essentiality profiling reveals gene...Karolinska Institutet, in Sweden. It can be very time consuming to design 5 guide RNAs (gRNAs) targeting... failing to upregulate the protein following stimulation would then be separated by sorting, and subsequently...much, showing that we are far from consensus on optimal gRNA design. With Green Listed you can also easily...PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize...

  14. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    .... Other molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific...chemical signals such as IL-8, IFNγ, and colony stimulating factor 1 (CSF-1). Additionally, tumor cells recruit...cell-surface proteome to inhibit expression of co-stimulatory molecules needed for T cell effector function... most acclaimed form of immunotherapy in recent times. These engineered T cells are capable of recognizing...immunotherapy targets can be utilized at the same time. A combination of tolerance marker blockade and ...

  15. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...antibodies through their HLDA workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has... Triton X-100; and methanol, we found that the optimal perm/fix solution depended on the specific antibody...wild-type and the KO population (Figure 1), further optimization can be performed to both maximize the separation...concentration (this can also save money!); and optimizing fixation/permeabilization reagents for intracellular...intracellular targets. Since knockout experiments can be time-consuming and expensive, it may be helpful to find...caused by siRNA is also transient, and as such, the optimal knockdown of the protein target will depend on ... is not efficient at the protein level at the timepoint studied Off-target activities of the RNAi Correctly...

  16. Promoters

    Type
    Guide
    ...RNA polymerase transcribes DNA to mRNA which is ultimately translated into a functional protein. Thus the...

  17. Chemogenetics Guide

    Type
    Guide
    ...G-protein and activate neuronal firing through stimulating phospholipase C, which releases intracellular...

  18. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...Addgene would also like to thank Viviana Gradinaru and Tim Shay for their support and input. Resources and ... Hudry et al, 2023; Hinderer et al, 2018). The optimal dose for each particular experiment is determined...systemic AAVs are a good fit for your experiment, it’s time to start selecting and testing systemic capsids!...

  19. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...receive different types of stimulation may help to refine and optimize your stimulation parameters as well. ...prefer to spend most of its time in the chamber in which it receives the stimulation (conversely, if the circuit... and stimulate them for 30min to an hour and observe what happens. You never know how stimulation of a...personally interested in include: intracranial self-stimulation (ICSS) and place preference. The lab I work in... electrode and non-specifically electrically stimulating an ill-defined region of the brain. With the ...particular hole (the active hole) induces laser stimulation of the chosen brain circuit, while a nose-poke...different patterns and the animal only receives stimulation in one of the two chambers (see image below)....

  20. Technical Design of a Western Blot

    Type
    Blog Post
    ...quickly Wet transfer Increased time Handcast gel Increased time; have to make as-needed Increase...costs; increased optimization Normalizing to total protein loading Increased time; increased reagents...from lot-to-lot, and have to be re-validated each time a new lot is purchased. Monoclonal antibodies, on...available for your target! Incubation temperature and time Incubating at a lower temperature reduces background...to reduce background while keeping the assay run time to two days.  Indirect or direct detection method...as needed. However, more antibodies mean more optimization and likely higher background noise as well, ... Good for higher weight proteins Long running time   It’s important to note that Bis and Tris gels...
Showing: 101 - 120 of 927 results