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We narrowed to 527 results for: HA tag

Showing: 141 - 160 of 527 results

  1. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...CDSns, coding sequence with no stop codon; TAG, C-terminal tag; TER, terminator. Created with BioRender....plasmid part? Do you want to fuse your protein to any tags or fluorescent markers? If you’re creating a multigene...all the learning and planning, and your MoClo kit has just arrived! MoClo kits from Addgene will most likely...further assemblies. This will help confirm each part has ligated in the right order and direction (which is...directly on the choices you make during the early stages of the process. Some questions to start with are...

  2. Hot Plasmids - November 2023

    Type
    Blog Post
    ...lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters, specific time points...plasmids here! Lin D., Li X., et al. (2023). Time-tagged ticker tapes for intracellular recordings. Nature...off-target effects of gene therapy, Feng Zhang’s lab has developed a system called DIRECTED (Delivery to Intended...on-target delivery with various delivery modalities and has the potential to reduce negative side effects. This... assays like IHC. In fact, Anti-PSD-95 [K28/43R] has been one of the hottest antibodies in Addgene’s collection... collection since we began distribution and has been verified to perform well in standard IHC and western...externally-timed bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record...

  3. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ... fusion tag to be inert, to not produce artifacts in your experiments. Considerable effort has gone into...solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid...world, is not inert. In all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently dimerize...dimerizing protein of interest fused to a dimerizing FP has potential to form large stable polymers of dimers... monomerized with an A206K mutation [2]. Addgene has many of these constructs and they are designated ...oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can be solved, in most cases, by using...

  4. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...this technology and its advantages? Fujii: In the last several years, my lab has been working on development...immunoprecipitation (ChIP) (iChIP) technology, which uses locus-tagging and affinity purification, in 2009. However, iChIP...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system...joined Addgene's Advisory Board.  Addgene: Your lab has worked extensively with enChIP systems. Can you describe... systems to Addgene in the near future. Addgene: Has your lab been working on any other technologies that...plasmids. The merit of depositing plasmids to Addgene has been spectacular. We've got more than one hundred...

  5. The Blue Flame Award: Celebrating Addgene's Most Requested Depositors

    Type
    Blog Post
    ...indicates that a plasmid has been requested 20+ times   A red flame means a plasmid has been requested 50+ ...love to see you show it off! To share your photo, tag @Addgene and #happydepositor on Twitter. All Blue... Castro of University of Michigan says, “Addgene has transformed the way in which we share plasmids and...making it easier, cost effective and faster; this has helped accelerate scientific discoveries.” Occasionally...+ times   A blue flame means a plasmid has been requested 100+ times Since we so sincerely appreciate ...have at least one plasmid deposited at Addgene that has been distributed more than 100 times. We hope these...sharing because, as UNC’s Bryan Roth says, Addgene has “literally enabled sharing with 100's of investigators...

  7. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    ...%}{% end_module_attribute %}{% module_attribute "tag" is_json="true" %}{% raw %}"module"{% endraw %}{%...Region – Let's Go! Plasmids for Endogenous Gene Tagging in Human Cells Resources on addgene.org Allen...have the biggest impact. We should know — Addgene has built its foundation on this idea! And we believe... cell populations and regions of the brain. This has culminated in a collection of AAV enhancer plasmids... to express exogenous genes — a feature the AIBS has capitalized on. Figure 1: Schematic of transcriptional...Allen Institute’s Genetic Tools Atlas (GTA). The GTA has a wealth of information and includes annotated images...target a very specific cell type. However, Addgene has partnered with the AIBS to showcase the “best-in-...

  8. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...cut, activate, interfere, visualize, nick, purify, tag, scaffold, CRISPR-display, and dCas9-FokI. This table...Alternatively, you can select a target sequence which has already been shown to work in CRISPR experiments....experiments. Addgene’s dedication to resource sharing has led us to develop a searchable and sortable datatable ... to the fact that every gRNA listed in the table has been used effectively in a published article. This...target sequence, and model system. Even if a gRNA has been demonstrated to work in your system, it is worth...that locus. The data in the validated gRNA table has been derived from scientists’ submissions, either...

  9. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...generate one molecule of GFP-tagged LC3 for every one molecule of RFP-tagged LC3∆G. Like the unmodified ...number of autophagosomes. This approach, however, has inherent limitations because it turns a dynamic process...

  10. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...Fluorescent Protein Should I Use? However, fusion proteins tag your protein of interest with a fluorescent reporter...protein. One downside of fusion proteins is that the tag can alter the structure of the protein. In some cases...timing of expression can also be affected by the tag.  Protein expression, measured using antibody conjugated...measured using visible light scatter The optical system has both visible and fluorescent light sources. Visible...autofluorescence from your target cell type or that has arisen from fixation. Also include an isotype control...secondary antibody control in which the primary antibody has been omitted from the staining protocol. This control...gating during data analysis.  We hope that this blog has provided you with an overview of flow cytometry for...

  11. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...BirA* biotinylation activity ceases. After biotin tagging of proteins in both BioID or Split-BioID, cells...depending on which terminus of the bait proteins were tagged but also which fragment of BirA*, either the N-...N- or the C-terminus, the bait proteins were tagged with. It’s best to test all combinations and pick ...opposed to an individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID,... of the fusion protein. Advantages of Split-BioID Split-BioID’s key advantage over other PPI methods, ...proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that it allows for the validation...

  12. Tips for arabidopsis transformation

    Type
    Blog Post
    ...terminally tagged fusion proteins with fluorescent (YFP, GFP, CFP, RFP, etc.) or affinity tags (HA, FLAG)....putative T1 transformants: When your T1 is on soil and has recovered from the stress of selection you can clip...cell fate maintenance in the stomatal lineage and has made countless transgenic plants in the process. ...

  13. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...rate. In many cases, however, a GFP-tagged cell line or organism has already been made, and it would be... Even though GFP has been so thoroughly characterized, it turns out this protein has a few more secrets...photoconvert GFP from green to RED! This simple technique has been shown to work in plant, Drosophila and mammalian...is great news for microscopy enthusiasts! Addgene has empty backbones containing GFP variants used by Sattarzadeh...

  14. A Look at Addgene's QC Process

    Type
    Blog Post
    ...plasmid they ordered contains the correct inserts, tags, promoters, and other critical functional components...on the information provided and vector type  Any tags, selectable markers, and fusion proteins listed ...including the propagation of plasmids in bacteria — has many points at which small errors can be introduced...Catching and addressing QC issues Addgene’s QC team has developed a process for identifying, categorizing...

  15. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...fusion protein The MLV Gag protein fused to a Flag-tagged SpCas9 is incorporated into Nanoblades, just like...insertions like the addition of a fluorophore or tag. To utilize HDR, a donor template containing the ...particles with polybrene, a cationic polymer. Polybrene has long been used to increase retroviral transduction...of mouse zygotes and mice with Nanoblades CRISPR has also been extensively used to create transgenic mice...

  16. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...measurable through an epitope tag such as FLAG/His/GFP. Fluorescent protein tags, which correlate closely ...different epitopes. Detection of overexpressed (tagged or untagged) protein When expression plasmids are available...expensive, it may be helpful to find an antibody that has already been validated for flow cytometry through...Characterization through Open Science) initiative has now started producing flow cytometry characterization...overexpression. Additionally, if the cell line used has endogenous levels of expression, this can make the...TIM-1 (Single pass type-1 membrane protein), which has been designated CD365 in the most recent HLDA workshop...as part of the YCharOS open science ecosystem. It has received in-kind support to produce this data from...

  17. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    ...after the first tagging step. As an additional benefit of plexWell™’s sequential tagging procedure, the...with our plasmid assembly pipeline. To date, this has resulted in high quality NGS-based assemblies for...

  18. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...controlled easier than formation of GST dimers. Avi-Tag plasmid for improved biotinylation selectivity A ...fusion protein from a bacterial extract. The MBP tag can be removed using Xa protease. The carboxyl-terminus... binding protein (MBP, New England BioLabs). MBP has a similar function as GST in that it provides a method...amino acid) AviTag sequence in a protein and adds biotin to a lysine residue within the AviTag. However,...carboxyl-terminus of the fusion protein includes an AviTag and a 6X HisTag for affinity purification (see plasmid ...dramatically increased accessibility of the AviTag and HisTag, which were “buried” in a few of our proteins...fusion proteins - but it would remove both the AviTag and HisTag. We were surprised that we couldn’t find ...

  19. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...fluorophore-tagged anti-rat IgG1 secondary to detect Protein A and green fluorophore-tagged anti-mouse ...imaging experiment   We hope that this blog post has answered all of your questions about the utility ...

  20. Choosing Your Fluorescent Proteins for Multi-Color Imaging

    Type
    Blog Post
    ...is the ability to follow multiple fluorescently tagged species simultaneously. To do so with fluorescent...generally the most detectable and so should be used to tag your least abundant proteins, with the blue and infrared...protein, if you know that your protein of interest has a similar spectrum. For example, here's a screenshot...#49004) to the spectra of both mCherry and TagRFP.  Here, the TagRFP spectrum is shown in the darker colors...peak of the TagRFP excitation and the emission filter collects a larger fraction of the TagRFP emission ...with this set are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T, and an infrared... emission. For this filter set, we would expect TagRFP to give a brighter signal than mCherry. In general...
Showing: 141 - 160 of 527 results