...   Myc1-9E10 and the Myc-tag today Since Munro and Pelham introduced the Myc tag to the world, it...commonly used Myc-tag. Who first started using it in protein tagging experiments? Why Myc? When did the...explore… the life and times of the myc tag.  Origins of protein tags For about as long as the disciplines... Find recombinant anti-c-Myc [9E10] at Addgene Find plasmids with the myc-tag  ...antibodies against the Myc tag have been generated in an attempt to overcome some of Myc1-9E10’s shortcomings...probe the functions of the human c-Myc gene (Evan, et al. 1985). Myc was one of the earliest identified...for protein tagging approaches” was not part of the goal when Evan’s team generated their Myc antibodies...

...guess that the Myc-tag is one of them (and not just because of the title!) The Myc-tag is a string of ...proto-oncogene c-Myc. Since the generation of the first anti-c-Myc [9E10] (Even, et al., 1985), the Myc tag has become...doi.org/10.1101/2021.09.17.460684.)   Myc-tag antibody anti-c-Myc [9E10] now available! by: Ashley Waldron...detecting protein tags.  And when you think of protein tags, what are the first tags that come to mind...of the most commonly used protein tags - a quick search for ‘Myc tag’ in the Addgene catalog pulled up... of Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the...Improved voltage indicator: JEDI-2P Novel class of light-gated potassium channelrhodopsins Myc-tag antibody...

...purification tag blog!).   Applications of epitope tags   The tags A number of epitope tags exist...monoclonal (M2) as well as a chimera (rabbit). Myc The Myc tag is a 10 amino acid peptide (EQKLISEEDL) derived...guidance on designing a GFP tag Addgene has you covered!   Potential tag issues Tags have a lot of major benefits...fix your antibody woes.   What are epitope tags? Epitope tags are short peptides introduced at the N or...system. Tagging can also aid in protein purification! In this blog, though, we are focusing on tagging for...amino acids on the C terminal of the tag. Of the most common tags, FLAG is the most charged, with 5 negatively...charged and 1 positively charged amino acids. Myc tags are frequently used for western blot, flow cytometry...

...Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built into a GFP scaffold. Some GFP... secondary antibody staining HA Tagged GFP, RFP, H2A2B Myc Tagged RFP, H2A2B Chicken IgY (Y) Labeled...RANbody is detectable by two different tags: a reporter and an HA tag. The reporter comes in three varieties...Spaghetti Monster tagged RANbodies can be detected either with a labeled anti- HA or -MYC antibody or with... Spaghetti Monster: HA (H) or Myc (M) versions Labeled anti-HA or -Myc antibody Staining cultured cells...lot-to-lot variability in activity. No Genetic Tagging - Since antibodies are produced in animals instead... via recombinant DNA, they can’t be genetically tagged and instead are conjugated to reporters (i.e. fluorescent...

...Why do we need new epitope tags? Short epitope tags like FLAG, HA, and Myc can adopt a number of different...the effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags. Impressively, IT6-GFP did not... looking to a tag a protein, inntags IT5 and IT6 are a great place to start. These tags are easy to use... about other Protein Tags in our Plasmids 101 Series. Get the Inntag Plasmids Tag endogenous proteins ...major caveat - a tag may interfere with protein localization and/or function, so each tagged protein must ... Aldea and collaborators have created a set of “innocuous tags” (inntags) less likely to alter a protein...proteins found that 20% of tagged proteins did not localize similarly to their untagged counterparts. These ...

...PCR based detection, DNA staining, fluorescent tagging, agar plating, etc. Be wise to choose any of the...you encounter mycoplasma contamination. Causes of mycoplasma contamination The mycoplasmas enter the cell...antibacterial drugs like penicillin and streptomycin. Mycoplasma can pass through filtration methods because...cell cultures are kept will increase the risk of contamination. Mycoplasmas can spread from these sources... growth of mycoplasmas. The serum is nutrient-rich and also the best source for mycoplasma proliferation...suppress the growth of mycoplasma but cannot eradicate it completely. When mycoplasmas are seen in the culture...highly expensive). Detecting mycoplasma contamination Detection of mycoplasma by naked eyes or optical microscope...

...which attaches a Myc-tag and a His6-tag to the end of each sybody candidate. These tags are used in a final...starts out with anti-Myc antibodies bound to an ELISA plate to capture Myc-tagged sybodies. Then, the ...pBXNPHM3 expression plasmids for the production of tag-free proteins for further characterization and analysis...Walter et al., 2020). No llamas required. The advantages of synthetic nanobodies Nanobodies are small ...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...step using ribosome display because it has the advantage of starting out with a larger initial starting...

...replacing C-MYC with L-MYC in the episomal reprogramming cocktail by Okita et al. increased the reprogramming...enhances iPSC generation as does replacing C-MYC with L-MYC [10, 11]. While the mechanisms underlying the...reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation of clinical grade hiPSCs,...that encoded six factors - OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large T antigen (SV40LT). The method...pCXLE-hSK 27078 SOX2 and KLF4 pCXLE-hUL 27080 L-MYC and LIN28 pCXLE-hOCT3/4-shp53-F 27077 OCT3/4 and...direct reprogramming by transformation-deficient Myc. Proc Natl Acad Sci U S A, 2010. 107(32): p. 14152...hiPSCs than age-matched fibroblasts [3]. These advantages make PBMCs the ideal source of somatic cells ...

... cleavage tags Frequently, a single tag is not enough. What if you need one tag to increase solubility...to fuse an affinity tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses...affinity tag is the polyhistidine (His) tag. Small and unlikely to affect function, His-tagged proteins...greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function Notes...to all tags, but rather a quick overview of why scientists use tags, with a few time-tested tags and fusion...protein. Unlike other epitope tags – which when doubled or tripled increase the tag size quickly – modifying...you generally don’t want more tag than protein), 2-3 tags is increasingly common. Tandem affinity purification...

...genome-wide. It was designed based on MYC-ChIP-seq data from several MYC-dependent cancer cell lines. It can...selection marker, Thy1.1, in addition to puromycin resistance.  The MYC-CRISPR library targets E-boxes genome-wide... can be applied for identification of essential MYC binding sites in a wide range of human cells.  A new...’s lab previously developed such a system in Streptomyces, a prokaryotic genus capable of producing a ...different biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The...oxytetracycline enzymes. Next, this system was used to recreate natural biosynthetic pathways. Biosynthetic pathways...Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline samples, were engineered...

...different types of tags, including epitope tags (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry... His-tagged protein purification work flow by nickel column.    Affinity tags vs. other tags There.... The MBP tag, although large, can actually increase expression and solubility of the tagged protein (...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...cleavage at sites immediately following the tag. Unlike MBP, the GST tag can be cleaved off from the fusion protein...

...proteins (GFP and YFP) and protein tags (c-MYC and HA). We have also created the Monoclonal Antibody Research... associated hybridoma cell lines. The DSHB was created by the National Institute for Child Health and ... which are each produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells ...individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing a ...partially humanized rabbit antibodies that exhibit increased avidity to high profile antigens; and 4) generating...

...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...insertion tagging), developed by the Hornung Lab, allows users to easily create C-terminal tag fusions ...between the tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable...genetic tag and marker sequence into the targeted loci. The selectable marker can be removed by Cre-mediated... inserting a tag at multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes...endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein Tagging. Beyond simply...

..., codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, ..., 6xHIS, MBP, Myc). These constructs include proteins like nonstructural proteins 1 to 16, spike proteins...expression constructs contain protein coding genes, tags, truncations, and codon optimizations similar to... downstream experiments and offer flexibility in tag, cleavage, codon usage, and expression system. Mammalian...expression vector are also available. These contain no tags and are intended to be used in the production of...sequencing by NGS. Addgene’s QC scientists immediately screen these plasmids to confirm they contain the expected...of plates will have select samples prepared and screened by NGS to spot check the samples, the rest will...

...antibody recognition of epitope tags. Thus they have created antigenic protein tags called “spaghetti monster...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 ...Viswanathan, 2015). The principal advantage of epitope tags for IHC is the availability of reliable primary... are unavailable entirely. The other advantage is that these tags are small (8-12 amino acids) and neither...composed of FPs fused to multiple epitope tags. Indeed, to create the smFPs Looger’s team strategically inserted...

...anti-FLAG tag antibodies By Ashley Waldron Of all the epitope tags we know and love, the FLAG tag (amino ...Endothelial-specific AAV delivery Recombinant anti-FLAG tag antibodies Peptide-assisted genome editing for ...Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and SV40 Nuclear Localization Signals, and GFP. B... purify or image a recombinant protein, the FLAG tag has proven itself a good option for essentially all...collection includes over 10,000 plasmids with a FLAG tag (!), but did you know that we distribute ready-to-use...  Neurogenetic alchemy: CAP-Mac unleashes the secrets of primate brains with brilliance By Hridaya Patel...authors used a multispecies approach to generate and screen a library of AAV9 variants (Chuapoco et al., 2023...

... viral particles that express oncogenes, such as Myc. Viral vectors also have the largest genomic footprint...transgenes is a concern as well, and reactivation of Myc or other oncogenes after differentiation has been...formation in iPSC-derived and iPSC-transplanted mice. Cre-deletable or Tet-inducible lentiviruses address some...considered safe for clinical applications. Excisable Cre-lox lentiviral vectors: This approach is the same...standard lentiviral vector, but transient expression of Cre recombinase allows for deletion of the inserted transgenes...including reporters and RNAi) for:  NANOG  OCT4 SOX2 MYC KLF4 LIN28 Check out these plasmids if you’re interested...due to the risk of insertional mutagenesis. Random integration also creates heterogeneous iPSC cell lines...

... been deposited with Addgene and include MYC- and FLAG-tagged TZAP constructs (wild-type and truncation...nonoverlapping emission spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and... transcription factors OCT3/4, SOX2, KLF4, and c-MYC is the established model for reprogramming somatic...instance, inserting PhoCl in between a protein and a tag, such as an NLS or NES, enables control over localization...light-dependent Gal4 transcriptional activation and Cre recombination. Find PhoCl plasmids at Addgene. Zhang... for easy removal of the resistance cassette via Cre recombination. Each knockout includes unique barcodes...gRNAs used for TZAP gene editing, and a Hi6xs-MBP tagged TZAPznf9-11constructs used for bacterial expression...

...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) ...crRNA and/or try out an alternative tag (3xFLAG, Myc, V5, or HA). Tag-specific PCR genotyping can tell if...endogenous gene tagging in human cells Plasmids 101: Protein Tags Fluorescent tagging of endogenous genes...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...

...dedifferentiation stage involves overexpression of four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that...More (due to additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming...transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox...lineage, without dedifferentiating into a pluripotent stage. Cellular reprogramming The reversal of a differentiated...reprogramming [1]. The process can be divided into two stages: Dedifferentiation - Conversion of adult somatic...differentiated somatic cell to revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate...reprogrammed to iPSCs successfully [10-12]. The next stage of reprogramming consists of redifferentiation of...