We narrowed to 526 results for: HA tag

... 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...

...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...

...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... domain than simply removing a repressor. Tet-On has since become one of the most popular methods to induce...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 ... Karpf 167935 pLenti-tetON-KRAB-dCas9-DHFR-EF1a-TagRFP-2A-tet3G Tet-inducible expression of KRAB-dCas9...

... 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...

...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...

... 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 ...

...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...

...KB Fujii iChIP/enChIP to purify genomic DNA FLAG tagged dCas9 PDF, 110 KB Goldstein Nematode: gRNA design...PDF, 109 KB Mendenhall and Myers Mammalian: FLAG tagging endogenous proteins pFETCh_Donor ; additional HDR...cloning in gRNA: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) PDF, 49 KB Vosshall and Matthews (Link opens...

...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...

...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...

...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...

...blog has you covered. Happy voltage sensing!   Figure 3: Components of the types of voltage indicators...approaches. Functionally, the voltage sensitivity of the available VSDs has been successful even at the ... Several adaptations of this method have been to tag, cage, uncage, and trap the chemical fluor with the...single neuron level in vitro. In vivo monitoring has also been achieved with these sensors, but further...research tools for studying voltage within a cell are genetically encoded voltage indicators (GEVIs) and chemigenetic...Types of Voltage Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive...occurring voltage indicator. To give an example, a genetically encoded phosphatase (voltage sensing) hydrolyzes...