...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...

..., 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...

...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...

...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...

... 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...

...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...

... 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...

...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...

...DNA for use with virus-mediated delivery has many advantages for research. Viruses facilitate the delivery...Kb 80-130 Dividing Yes Moderate * Recombinant AAV has a low frequency of target cell genome integration...

...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...

... 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...

...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...

...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 ...

...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...

... 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 ...

...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...

...AAV2 capsid with the N587 residue replaces with a TAG stop codon Mali 130878 pAAV-DJ-N589X AAV-DJ-N589X...-DJ capsid with the N589 residue replaced with a TAG stop codon Mali 166921 AAV-F in pAR-9 AAV-F AAV rep...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...

...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...

...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...

...Research Cancer Research Plasmids and Resources Addgene has a large variety of resources for use in cancer signaling...vectors for in vitro and in vivo research. TP53 Mutagenesis Library : Pooled library from the Hahn and Root...