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We narrowed to 520 results for: HA tag

Showing: 31 - 60 of 520 results

  1. Plasmids 101: Degron Tags

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    Blog Post
    ...degron tags. What is a degron tag and why should I use one?   Degron tags are a class of protein tag which...amenable to being tagged, e.g. tagging some proteins can affect their function, the SMASh tag self-cleaves ...). Choosing a degron tag Are all tags created equal? No! Choosing the ‘right’ tag does depend a lot on...Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation tag (dTAG) system functions...degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and the endogenous cellular...machinery to function. The dTAG is the smallest tag reviewed here, in regard to total tag size, which can also...

  2. Adeno-associated virus (AAV) Plasmids

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    Collection
    ...Plasmid Gene/Insert Vector Type Backbone Mutations Tags PI Do you have suggestions for other plasmids that...products that mediate AAV replication. This requirement has been circumvented with “helper-virus free systems... contaminants of adenovirus stocks. One major advantage of using AAV for research is that it is replication-limited...

  3. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...strep-tag and, more recently, OLLAS (Escherichia coli OmpF linker and mouse langerin) and Sun Tag (Viswanathan...antibodies can have low affinity for these tags and even multimeric tags are frequently insufficient for detection...FP and epitope tags, researchers in Loren Looger’s lab have developed new molecular tags combining the ...antibody recognition of epitope tags. Thus they have created antigenic protein tags called “spaghetti monster...experiments with tag-specific antibodies. The most famous ones are influenza hemagglutinin (HA), myelocytomatosis...With the limitations of epitope tags and fluorescent proteins, it has been crucial to develop new tracers...provided better labels than fusions to 3 or more HA tags. 4. High-resolutions microscopy As indicated above...

  4. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed... scarless tag insertion.   In addition to the obvious advantages SapTrap offers for tagging a single genetic...between the tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable... inserting a tag at multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes...insertion tagging), developed by the Hornung Lab, allows users to easily create C-terminal tag fusions ...endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein Tagging. Beyond simply...expression levels. CRISPR tagging systems offer users an easy and efficient way to tag endogenous proteins ...

  5. Antibody Guide

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    Collection
    ...directly to the sdAb, or by using a secondary against a tag, such as His, incorporated in the sdAb. sdAbs can... immune response to it. Once the immune response has developed, the antibodies are collected, tested against...epitopes on constant regions of other antibodies. This has useful research applications, and is covered in the...exist as monomers. Each antibody class therefore has a valency, or number of binding sites available per...of antibodies multiplied by two, as each antibody has two binding sites. Isotype Description Configuration... must be able to detect if and where an antibody has bound. Signaling molecules , proteins that emit a...to avoid confusing data. Check that your machine has appropriate lasers and filters for each fluorophore...

  6. Tetracycline Inducible Expression

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    Collection
    ...expression gene of interest fused with 3xFLAG and 2xStrep-tag from TRE3G promoter for Tandem-Affinity Purification...basic tool to control gene expression, the system has been engineered with a variety of improvements. A...promoter. And, most importantly, the TetR protein has been modified to provide two classes of tet-responsive...TetR and derivatives like rTetR, rtTA, and tTA. Dox has good tissue distribution, low toxicity, a known half-life...operator ( tet O, a 19 nucleotide sequence, TCCCTATCAGTGATAGAGA). In normal conditions, the TetR protein ... (Tet-On) In 1995, Gossen et al. used random mutagenesis to identify the amino acid residues of TetR that... Karpf 167935 pLenti-tetON-KRAB-dCas9-DHFR-EF1a-TagRFP-2A-tet3G Tet-inducible expression of KRAB-dCas9...

  7. Plasmids for Stem Cell Research

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    Collection
    ...related to embryonic stem cell use and has the major advantage that iPSCs can potentially be derived from... be used to create cell lines with endogenously-tagged gene variants or browse Addgene’s entire collection...Capecchi Replicating EBNA1 episome Human Fluorescent-tagged EBNA1-mediated expression of human Oct3/4, shp53...dosage tracking of reprogramming factors Fluorescent tagged episomals for stoichiometric induced pluripotent...development. Unlike other cell types, each stem cell has the potential to either remain a stem cell or become...can be faster, bypasses intermediate pluripotent stages, and can occur without cell division, all of which...mutation and minimize tumorigenicity, an inherent disadvantage of iPS cell technology. Browse Addgene's collection...

  8. Antibodies 101: Affinity Tags

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    Blog Post
    ... His-tagged protein purification work flow by nickel column.    Affinity tags vs. other tags There...different types of tags, including epitope tags (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry...GST antibody? Addgene has you covered! His His tags, often called polyhistidine tags, typically include ...planning, affinity tags can make protein purification a cinch. Types of affinity tags, the proteins they...are all reviewed here. What are affinity tags? Affinity tags are used primarily as protein purification... common tags available. Maltose binding protein Maltose binding protein (MBP) is a 45 kDa tag which can.... The MBP tag, although large, can actually increase expression and solubility of the tagged protein (...

  9. Tag Your Favorite Yeast Genes with Ease

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    Blog Post
    ..., modular plasmids, combining fluorescent tags, epitope tags, protease sites, and selection markers. These...fluorophore tagging vectors for your single or multi-color imaging experiments. Interested in epitope tags? Others...have genome tagging systems enabled you to advance your research? Do you have a favorite tagging system that...at the University of Utah, has built a complete collection of yeast tagging modules with each possible...linker-(epitope tag)-ADH1 terminator-(selection marker). Additionally, Dr. Formosa has deposited six plasmids...Researchers have long taken advantage of this natural process to integrate protein tags into the genomes of S...especially useful in protein complex studies where tagging of multiple protein products is desired, as multiple...

  10. Hot Plasmids - October 2022

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    Blog Post
    ...the Myc tag has become one of the most commonly used protein tags - a quick search for ‘Myc tag’ in the...detecting protein tags.  And when you think of protein tags, what are the first tags that come to mind...guess that the Myc-tag is one of them (and not just because of the title!) The Myc-tag is a string of 10...channelrhodopsins. HcKCR1 has a maximum absorption of 540 nm and two-photon excitation around 1,040 nm, has larger photocurrents...Improved voltage indicator: JEDI-2P Novel class of light-gated potassium channelrhodopsins Myc-tag antibody...https://doi.org/10.1101/2021.09.17.460684.)   Myc-tag antibody anti-c-Myc [9E10] now available! by: Ashley... 3: Image of Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926...

  11. Hot Plasmids: Fall 2024

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    Blog Post
    ...epitope (GKPIPNPLLGLDST) has been used as a tag since 1992 (Hanke et al., 1992), and has proven itself a versatile...Epitope tag antibody collection grows By Ashley Waldron We are excited to highlight the Anti-V5 tag antibody...immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen. J Gen Virol., ...Figure 3: Expression of fluorophore-tagged GOLG-4 through eight stages of embryonic development. Representative...find in this post: Anti-V5 and other new epitope tag antibodies from IPI New tool for structural studies...studies of E3 ligase cereblon Optimized mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene...Innovation (IPI), part of a new collection of epitope tag antibodies available through Addgene. Anti-V5 [IPI-SV5...

  12. To Each HIS Own

    Type
    Blog Post
    ... on the polyhistidine tag. What is the polyhistidine tag? The polyhistidine tag is a string of 6 or more...polyhistidine tag, alternative fusion tags were used, such as S. aureus’ Protein A (Moks 1987). These tags, however...conditions.   Find polyhistidine tag plasmids here! Where should I place the tag and how many do I need? When...containing a polyhistidine tag, you’ll need to decide whether to add the tag to the N- or C-terminus and...requires the tag to be accessible to the metal ions once the protein is folded. Sometimes a tag placed on... however, tag placement can affect protein function. B, The string of histidines in the tag is typically... small size of the polyhistidine tag minimizes the chance of the tag disrupting normal protein function...

  13. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...epitopes tags such as HA and MYC in vivo. The authors found that fusions to one particular tag, the IT6... for epitope tagging: Inntags When fused to your protein of interest (POI), epitope tags give you the...production of a new series of epitope tags, inntags (innocuous tags), that they believe will have fewer ...expressed either without a tag or as multiple fusions with different N- and C-terminal tags. Additionally, by ...expression of fusions between the POI and a TAP tag (a tag with two epitopes that can be used to sequentially...two dimeric Broccoli tags (2xdBroccoli) to further enhance fluorescence of the tagged RNA. Bacterial and...containing a variety of N- and C- terminal tags. The Santalucía Lab has developed a cloning toolkit based on...

  14. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... the best outcomes. The go-to in vivo option AAV has become the preferred delivery method for in vivo ... other viral vector delivery methods. Lentivirus has limited packaging options, with most lentiviral vectors...possibilities when designing screens. Each serotype has its own tropism, or a preference to infect specific...BioRender.com. Beyond the identified serotypes, AAV also has psuedotypes. Psuedotyping is the process of making...Lentiviruses have capacities of 8–12 kb, while AAV only has approximately 4.7 kb — which means that when using...the genetic landscape of tissue-specific diseases has grown exponentially, aided by advancements in in ...ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in vitro, ex vivo CRISPR screens...

  15. Fluorescent Protein Guide: Subcellular Localization

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    Collection
    ...Plasmids encoding fluorescent proteins tagged with genes or peptides with known subcellular localization... protein of interest with known markers. Addgene has assembled a collection of fluorescent fusion proteins...protein of interest with a fluorescent protein that has a different emission wavelength. For example, if ...mScarlet-I Dorus Gadella 50057 pLYS1-FLAG-MitoGFP-HA Mitochondria MCU GFP Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer...Voeltz 79802 pTag-RFP-C-h-Rab5a-c-Myc Early endosomes Rab5a TagRFP James Johnson 79801 pTag-BFP-C-h-Rab5a-c-Myc...Mothes 79806 pTag-RFP-C-h-Rab11a-c-Myc Recycling endosomes Rab11a TagRFP James Johnson 79805 pTag-BFP-C-h-...endosomes Rab11a TagBFP James Johnson 79800 pTag-RFP-C-h-Rab4a-c-Myc Recycling endosomes Rab4a TagRFP James Johnson...

  16. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

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    Blog Post
    ...is to tag the protein of interest with a small biochemical “epitope tag” such that these tagged proteins... the tagging efficiency in the transfected cells, we count the number of tag-positive cells (tag ICC) ...endogenous gene tagging in human cells Plasmids 101: Protein Tags Fluorescent tagging of endogenous genes...crRNA and/or try out an alternative tag (3xFLAG, Myc, V5, or HA). Tag-specific PCR genotyping can tell if...each with a different tagged protein of interest, and a universal all-purpose “tag antibody” can be used...projects that require epitope tagging of a large cohort of genes (e.g. tagging genes representing a protein...pipeline for epitope tagging. Using our 96-format pipeline, we can routinely achieve 5-30% tag knock-in efficiency...

  17. Isolating Genomic Regions of Interest with the CRISPR System

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    Blog Post
    ...3xFLAG and biotin tags, use of tag systems including Protein A, 1xFLAG, 2xAM, and HA have been reported...2013). However, the fusion of dCas9 with a protein tag allows for the isolation of a genomic region of interest...cells by expressing dCas9 with an N-terminal 3xFLAG tag (3xFLAG-dCas9) and a gRNA targeting their genomic...Immunoprecipitation, using an antibody targeting the 3xFLAG tag, allowed these researchers to isolate dCas9 with ...chromatin or DNA fragment (Fujita et al., 2016). 3xFLAG-tag fused to dCas9, along with a biotinylated gRNA, were...were successfully used in this "in vitro" form. Any tag system can be used to isolate genomic regions of ...This system utilizes dCas9 fused to a biotinylation tag expressed on a plasmid, allowing for in vivo biotinylation...

  18. Bacterial Expression Systems

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    Collection
    ...purification. Commonly used tags, cleavage sites, and signal peptides include: Epitope tags: 6xHis, Flag, Strep...bacteria with different purification tag combinations, including PelB tags for localization to the periplasm...Protein Expression Protein Expression Epitope Tags Protein Tags Return to top Protein Visualization To determine...Strep II, c-Myc, HA, V5, GST Solubility tags: MBP, SUMO, TrxA, Mocr, NusA Cleavage sites: TEV protease, factor..., proteins are often expressed alongside epitope tags, which can also later be used for protein detection...removed using protease cleavage sites. Other protein tags and signal peptides are used to enhance protein ...localization: PelB, MalE, OmpA ID Plasmid Promoter Tags PI Additional Addgene Protein Purification Resources...

  19. Hot Plasmids - February 2022

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    Blog Post
    ... small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit multiple...SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag. ...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs ...b) Schematic of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of...cassette is advantageous to Cas9’s larger expression requirements (Figure 1), but Cas12a has generally .... The proteins, which are purified via histidine tags, are optimized for SDS-PAGE gels and can be detected...targets by: Mike Lacy Adding to their set of anti-HA frankenbodies (Zhao et al. 2019), Tim Stasevich and...

  20. Of Myc and Men

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    Blog Post
    ...introduced the Myc tag to the world, it has become one of the most commonly used tags in molecular biology...instance, the commonly used Myc-tag. Who first started using it in protein tagging experiments? Why Myc? When...explore… the life and times of the myc tag.  Origins of protein tags For about as long as the disciplines...was within the tagged protein rather than at the C-terminus. These features made this tag easier to use...“equi-kli-seed-le”). (3) Tag all the things!   Myc1-9E10 and the Myc-tag today Since Munro and Pelham... amino acids, meaning the tag could be quite small. However, substance P has an amide group added to its...high quality antibodies available - aka peptide tagging.  Key to this strategy was the existence of high...

  21. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

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    Blog Post
    ...join one peptide tag to another peptide tag. Addgene: What is the advantage of peptide tags for building ...locking two peptide tags together. SpyLigase overcomes limitations in the use of peptide tags, which often form...Howarth: Peptide tags can be very generic; you should be able to use the same peptide tag for all sorts ...Natl Acad Sci U S A. 2014 Mar 17. PubMed) Peptide tag forming a rapid covalent bond to a protein, through...form only weak and reversible bonds. Howarth’s team has already demonstrated in their PNAS paper that SpyLigase...discovered the domain. It is small, highly stable and has this amazing chemistry locking the domain together...cancer in particular. They recently introduced the SpyTag/SpyCatcher system, based on a protein isolated ...

  22. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

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    Blog Post
    ...system for protein tagging. We spoke with Mendenhall and Myers about their new CRISPR tagging system, which...www.addgene.org/crispr/tagging. Addgene: What motivated you to adapt CRISPR for use in protein tagging? Myers: Our...epitope tag onto the transcription factors. By the way, you can actually use this for tagging any protein...population of cells with the tagged transcription factor. The FLAG antibody has been well characterized and...factors. Our groups got together to adapt CRISPR for tagging experiments and it’s already looking like it’s ...correctly, the second plasmid inserts a small epitope tag called FLAG at the end of the transcription factor...of advice for people who may want to apply this tagging system to their own work? Mendenhall: We’ve made...

  23. Your Top Requested Plasmid in 2016!

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    Blog Post
    ... SpCas9 is also fused to a 3X FLAG tag at its N-terminus. This tag can be used to test for SpCas9 expression...Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9 and further variants ...modified version of CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines. Cas9...effective in certain cell lines. This point mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro (PX459...that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction sites ...

  24. Hot Plasmids: Winter 2025

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    Blog Post
    ...Twelvetrees Lab took advantage of the self-labeling tags SNAPf and CLIPf. These tags rely on synthetic dye...single- and double-tagged constructs (including "CLAP", combining an N-terminal CLIPf tag and a C-terminal...studying mEPSPs has been possible with conventional electrophysiological methods, it has remained difficult...dicots Self-labeling tags for single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors...2024.02.11.579803).  CLIP, CLAP, SNAP! Self-labeling tags for single-molecule biophysics By Mike Lacy In order...C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression and labeling approach ... The team deposited an array of constructs for tagging the kinesins KIF5A and KIF5B as well as backbones...

  25. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...protein available – you can always add an epitope tag or a fusion protein. Mammalian vectors One of our...to be a more robust expression system and has its advantages; however, remember that you will need accessory...pBAD LIC cloning vector (8A)) and have an epitope tag (6xHis or GST) to affinity purify the expressed protein...popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following...& Other Plasmid Protocols List of common epitope tags ...You will also have to make sure that your plasmid has been incorporated into the host organism, usually..., c-Flag pcDNA3, comes from the pcDNA series. It has the CMV promoter for mammalian expression, a multiple...

  26. 27 Hot Plasmids from 2016

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    Blog Post
    ...interactions. The Roux lab has deposited HA tagged BioID2 for N-terminal fusions and Myc tagged BioID2 for C-terminal...for an individual tagging event. Commonly used sequences, including fluorescent tags, a floxed Cbr-unc...proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly ...organisms has limited routine use of this technology. To streamline CRISPR/Cas9-based genetic tag insertion...generation lentiviral transfer vector. Each gene has been FLAG-tagged for convenient experimental use. The plasmids...with SpyTag/SpyCatcher and SnoopTag/SnoopCatcher The development of recombinant DNA technology has allowed...119 selectable marker, and “connectors” that link tags to the targeted gene are supplied to the reaction...

  27. Easing the Protein Purification Process with pCri

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    Blog Post
    ...of interest is purified using an affinity tag. Additional tags may be used to improve protein stability...researcher to test multiple tags for one protein quickly and easily. These tagged plasmids also include cleavage...remove the tag post-purification. A sample of pCri constructs: pCri-1a, -4a and -9a. 6xHis tags enable affinity...remove these tags from the purified protein. For more detailed information on these and other tags used in ...integral-membrane protein constructs) protein tag. MISTIC drags tagged proteins into the membrane and may improve...which includes plasmids with different varieties of tags, both N- and C-terminal. Each set of plasmids uses...solubility and stability. TRX and signal peptide tags help proteins with disulfide bonds fold properly...

  28. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

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    Protocol
    ... a Multiple Cloning Site (MCS) Adding short tags - ex: HA or Flag Cloning shRNAs Background For the purposes...technique can be used just as easily to add short tags or shRNAs to any vector (the procedure will simply... design). Let's assume that your favorite vector has a relatively limited MCS (BamHI - EcoRI - SalI) and...

  29. Visualizing Protein Turnover In Situ

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    Blog Post
    ...protein tags can be used to facilitate the visualization of specific proteins in cells. Tagged protein...containing AHA to a fluorescent alkyne tag. The newly synthesized and tagged proteins can then be observed with...the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative is the...here, there is the question of whether or not the tags influence behavior. A conference and a breakthrough... known as fluorescence noncanonical amino acid tagging (FUNCAT, Dieterich et al. Nature Neuroscience, ...protein of interest that also had incorporated the AHA tag after pulse labeling of a determined period of time...used for cell labeling Check out other protein tagging strategies Additional Resources on Addgene.org...

  30. What's New in CRISPR - Winter 2018

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    Blog Post
    ...Nature Methods Find the anti-CRISPR plasmids Tagging human transcription factors with GFP Kevin White...lab recently deposited a set of CRISPR plasmids to tag human transcription factors with GFP. As part of ...more about ENCODE Find the transcription factor tagging plasmids If you have a new CRISPR tool you’ve...dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras with higher...been many Cas nucleases developed for plants, it has been difficult to compare the performance of different...recently discovered in 2017. Now Dominik Niopek’s lab has adapted anti-CRISPR proteins for optogenetic control...
Showing: 31 - 60 of 520 results