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We narrowed to 117 results for: PEP

Showing: 1 - 50 of 117 results

  1. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

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    Blog Post
    ...just 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 ...assemblies? Howarth: Peptide tags can be very generic; you should be able to use the same peptide tag for all ...thanks for the T-shirt! References: SpyLigase peptide-peptide ligation polymerizes affibodies to enhance ...itself together, to produce irreversible protein-peptide interactions. In a study published in Proceedings...Proc Natl Acad Sci U S A. 2014 Mar 17. PubMed) Peptide tag forming a rapid covalent bond to a protein,...

  5. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...adapted "self-cleaving" 2A peptides into their muliticistronic vectors. These peptides, first discovered in ... these peptides are thought to function by making the ribosome skip the synthesis of a peptide bond at...cleavage with some of these peptides, no consensus has been reached on which peptide works best. Likely the...the specific genes cloned into the vector.3  2A Peptides To overcome some of the disadvantages of the IRES...between the end of the 2A sequence and the next peptide downstream.4 The "cleavage" occurs between the ...Proline.  The table below lists the four common 2A peptides employed by scientists. 2A cleavage is universal...the choice of specific 2A peptide will ultimately depend on a number of factors such as cell type or experimental...

  6. Of Myc and Men

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    Blog Post
    ...defined peptides. Munro and Pelham’s first published peptide tagging experiment involved a peptide from ...with unrelated peptides that already had high quality antibodies available - aka peptide tagging.  Key ...bound to the mid-protein peptide, while three bound to the C-terminal peptide. One of these C-terminal...from the neuropeptide substance P. This protein had a highly specific antibody to the C-terminal five amino...critical for antibody recognition. As such, the peptide could only be added to the C-terminus of proteins... tumorigenesis.  Using two different synthetic peptides corresponding to linear sequences within the middle...antibodies and identified Myc1-9E10 and its target peptide as a suitable pair for protein tagging. In two ...

  7. Hot Plasmids February 2024

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    Blog Post
    ...with Peptide-Assisted Genome Editing (PAGE). They fused the Cas enzyme to cell-penetrating peptides and...articles.  Here's what you'll find in this post: Peptide-assisted genome editing for primary cells Monomeric...delivery Recombinant anti-FLAG tag antibodies Peptide-assisted genome editing for primary cells By Susanna...and co-incubate it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with...reporter (mChe), with co-incubation with assist peptide (AP). C) Workflow for Cas-RNP-PAGE editing of primary... cells, by co-incubation of Cas-RNP and assist peptide. Adapted from Zhang et al. 2023 with permission...engineering of human and mouse primary cells using peptide-assisted genome editing. Nature Biotechnology, ...

  8. Plasmids 101: Gram Negative and Positive Bacteria

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    Blog Post
    ...bacteria, or monoderms, have a single thick peptidoglycan membrane. Common gram positive bacteria include...negative bacteria, or diderms, have a thinner peptidoglycan membrane and an outer lipid membrane. Common...salmonella and E. coli. Because of their thicker peptidoglycan layer, gram positive bacteria trap the crystal...violet (CV) is water soluble and can enter the peptidoglycan layer of both bacteria types. In water, the ...with the CV+, creating a complex within the peptidoglycan layer, staining the cell purple. When the alcohol...alcohol is added, it dehydrates the peptidoglycan layer, trapping the large dye complex created by the...the outer lipid membrane, and the thin inner peptidoglycan membrane is unable to retain the stain, leading...

  9. 22 Hot Plasmid Technologies from 2014

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    Blog Post
    ...for protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking. The...One 2013 Oct 7;8(10):e76117. Irreversible peptide-peptide ligation using SpyLigase Building off their...that promotes the formation of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated... demonstrated the use of the SpyLigase peptide-peptide interaction to link affibodies or antibodies against...degradation. Chemical approaches may require expensive peptides or small molecules and exceed a physiologically...

  10. Plasmids 101: Blue-white Screening

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    Blog Post
    ...encoding this section of amino acids (called the α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements...engineered a multiple cloning site (MCS) into the α-peptide (represented as an orange wedge in the figure on... your DNA (red) is cloned into this MCS, the α-peptide gets interrupted as shown in cell A, and thus will... An unsuccessful cloning reaction leaves the α-peptide intact, and therefore the cell will have a functional...insert. Any cloning artifact that disrupts the α-peptide DNA will also lead to a white colony. And also...disrupting the lethality. This is similar to α-peptide DNA disruption in the blue-white screen. Antibiotic...

  11. Antibodies 101: Epitope Tags

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    Blog Post
    ... What are epitope tags? Epitope tags are short peptides introduced at the N or C terminus of a protein...purification or detection of the tagged protein. These peptides consist of as little as 8 or as many as several... (rabbit). Myc The Myc tag is a 10 amino acid peptide (EQKLISEEDL) derived from the protooncogene of ...these species. HA The HA tag is a 9 amino acid peptide (YPYDVPDYA) derived from human influenza hemagglutinin... chicken) to this tag! GFP  GFP is more than a peptide – it’s a 238 amino acid protein that can also be...

  12. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...this kit have been used to purify a variety of peptides including antibiotic efflux pumps and ankyrin ...followed by leader sequences (encoding small leader peptides) downstream of which you can clone your gene of...interest. Translation of the standardized leader peptide prevents secondary structures in the mRNA encoding...from resulting in the production of a downstream peptide, the transcription terminators in this series are...for cloning. Furthermore, some plasmids contain peptides to facilitate protein folding and/or protease ...

  13. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...with peptide/protein analyses using ELISA. Peptides 72:4–15 . https://doi.org/10.1016/j.peptides.2015.04.012...state while a western blot detects denatured polypeptide chains. The protein regions available for antibody...between a protein in its native state versus the polypeptide chain. Consequently, antibodies validated for...

  14. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ... to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables selection...plasmid post transfection. The 2A "self cleaving" peptide causes the ribosome to translate Cas9-2A and the...the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone. This particular...cleavage site). Finally, in addition to the 2A peptide on its C-terminus, SpCas9 is also fused to a 3X...

  15. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

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    Blog Post
    ..., CRY2clust, was created by adding a 9-residue peptide to the C-terminus of human codon-optimized  cryptochrome...conditions. With its additional 9 amino acid peptide, the CRY2clust domain enables fast and reversible...original version containing CRY2 without the extra peptide. CRY2clust should prove a useful tool to further...Dronpa flanked a PKA substrate fused to the phosphopeptide binding domain FHA. The FHA1 domain binds to...

  16. Plasmids 101: Protein tags

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    Blog Post
    ... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the...protein. Though there are a number of chemical and peptide-based proteosome inhibitors, glutathione S-transferase...bacterially derived in-vivo biotinylation signaling peptide (Bio), flanked by hexahistidine motifs (6xHis) ... a combo tag comprised of a calmodulin binding peptide (CBP), a TEV cleavage site (more on that in a moment...

  17. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...overview of peptide synthesis. A ribosome interacts with mRNA and charged tRNAs to build a peptide. Image ...where they are incorporated into the growing polypeptide chain. One part of the tRNA contains the anticodon... codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...

  18. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ...others. Another peptide-based strategy uses an enzyme to covalently label the peptide with a fluorophore...proteins as described above. The smallest tags are peptides that are engineered to bind specifically and with...PubMed PMID: 10748019.  8. Lotze, Jonathan, et al. "Peptide-tags for site-specific protein labelling in vitro...

  19. Live and Let Dye: Self-Labeling Protein Tags

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    Blog Post
    ...1998 (Griffin et al., 1998), this 15 amino acid peptide contains a cluster of four cysteines (CCXXCC, where...using engineered ligases to attach a ligand to a peptide tag on your protein of interest or using non-natural...provide better labeling specificity than short peptide tags like tetracysteine (Jing & Cornish, 2011).... protein is functional. In some cases, a small peptide tag like tetracysteine might be the only option...

  20. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...to a target polypeptide. Multimerization in the SunTag system occurs via antibody-peptide labeling; specifically...specifically, cognate peptide epitopes fused to the protein of interest are recognized and thus fluorescently...designed photoswitchable dimers by fusing the SsrA peptide with the light-sensitive LOV2 domain of Avena Sativa...Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the asLOV2 domain and has reduced affinity...reporter proteins fused to the streptavidin-binding peptide (SBP). The addition of biotin disrupts this interaction...

  21. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ...synthesis Bactericidal 100 ug/mL Bleomycin glycopeptide Induces DNA breaks Bactericidal 5-100 ug/mL...Bacteriostatic 50-100 ug/mL in EtOH Polymyxin B polypeptide Alters outer membrane permeability Bactericidal...Chloramphenicol   Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH...

  22. Hot Plasmids - May 2022

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    Blog Post
    ...subcellular localization peptide. By combining three different localization peptides with four different fluorescent...mixed-subclone system using mStrawberry, localization peptides, and reporters that drive the expression of the...

  24. Antibodies 101: Conjugation

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    ...amino acids or peptide tags. 5) Strong protein-protein interactions with other peptides. Created with ...strong protein-protein interactions with other peptides, like an Fc-binding domain (Figure 1).  ...

  25. Antibodies 101: Affinity Reagents

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    Blog Post
    ...light chain regions, connected by a short linker peptide. The lack of constant region makes them less immunogenic...proteins formed by two Fab fragments bound by short peptide linkers. They are bispecific, with a lot of flexibility...a wide variety of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells. They...

  26. Illuminating Epigenetics with A FRET Based Biosensor

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    Blog Post
    ...consists of one substrate (the histone-derived peptide) which can localise the construct specifically ..., which binds selectively to lysine-methylated peptides. This construct is in the middle of a FRET pair...When methylation occurs at the histone-derived peptide, the methyllysine binding domain causes a structural...

  27. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ... DNA sequence to encode your peptide of interest and produce that peptide in your organism of choice. ...as easy as it sounds. Even for relatively short peptides, there can be many possible ways to encode them...

  28. Hot Plasmids - August 2020

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    Blog Post
    ... via BirA and AviTAG. pMAK contains the signal peptide of MBP, followed by the desired nanobody (NB) sequence...sequence, an AviTAG, and 6xHis tag. The MBP signal peptide allows the NB to be exported to the periplasm of...Protein Expr Purif, 2020. https://doi.org/10.1016/j.pep.2020.105607 A new multiplexed CRISPRi and CRISPRa...

  30. Plasmids 101: Biotinylation

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    Blog Post
    ... biology and biotechnology purposes.  Once a polypeptide is biotinylated (more on this below) it can be...Northern blotting Cell surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated...commonly used this method to attach proteins or polypeptides to a solid substrate (Holden and Cremer, 2003...

  31. Delivery Methods for Generating iPSCs

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    Blog Post
    ... them with peptides that help mediate their transduction, such as the polyarginine peptide described in...iPSCs using a ~14.5kb minicircle expressing a 2A peptide polycistronic cassette comprised of OCT4, SOX2,...

  32. Easing the Protein Purification Process with pCri

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    Blog Post
    ...protein solubility and stability. TRX and signal peptide tags help proteins with disulfide bonds fold properly...designed for these systems may include a signal peptide (SP) that routes the protein through the secretory...

  33. 5 Great Apps for Lab Life

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    Blog Post
    ...explain the properties of or draw an amino acid or peptide for some reason. It contains all twenty amino acids...allow you to write your colleagues’ names in a polypeptide chain, except if their names have a B, J, X, ...

  34. Plasmids 101: Mammalian Vectors

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    Blog Post
    ..., NIH3T3, CHO, 293HEK, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation 100...hygB HeLa, NIH3T3, CHO, Jurkat T cells Blocks polypeptide synthesis at 80S; inhibits chain elongation. ...

  35. Fluorescent Proteins 101: Luciferases

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    Blog Post
    ...beetle (Pyrophorus plagiophthalamus), deep-sea copepod (Gaussia princeps), Japanese firefly (Luciola cruciata...or naphthyl-luciferin analogs ATP No Deep sea copepod (Gaussia princeps) Natural 480 20 Coelenterazine...

  36. Antibodies 101: Fab Fragments

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    Blog Post
    ...IgG antibody (Figure 1): it is composed of four peptide chains (two identical heavy chains and two identical...recombinantly and engineered to be joined by a linker peptide, resulting in a product called a single-chain Fragment...

  37. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Millipore Sigma 12072 Antipain, Millipore Sigma 10791 Leupeptin, Millipore Sigma L2884 Aprotinin saline solution...month. 1000X protease inhibitor cocktail 25 mg Leupeptin 50 mg Antipain 250 mg Benzamidine 25 mL Aprotinin...

  39. Plasmids 101: Protein Expression

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    Blog Post
    ... can then be translated at the ribosome into polypeptide chains that make up the primary structure of ...recombinant proteins with a naturally secreted plant peptide allows for easier access and purification of a ...

  40. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ...short genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure...covalent attachment of a fluorophore ligand to a peptide tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine...

  42. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...neuronal circuits. Epitope tags are short antigenic peptide sequences, attached to a protein of interest (POI...simian virus 5-derived epitope (V5), the synthetic peptide FLAG, the synthetic streptavidin-binding strep-...

  43. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...

  44. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ...recombinant antibody. They are ~25 kDa single polypeptides that contain the variable light chain (VL) and... two chains are connected by a flexible linker peptide that is usually 15-20 amino acids long and made...

  45. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...extension and translation of the pSer-incorporated polypeptide. These three components were shown to effectively...proportional to a single phosphate moiety on the peptide of interest.  After confirming pSer incorporation...

  46. 27 Hot Plasmids from 2016

    Type
    Blog Post
    .... Mark Howarth’s lab recently engineered a new peptide/protein pair, SnoopTag/SnoopCatcher, that can be...link desired proteins together via a spontaneous isopeptide bond. When used in combination with their previously...

  49. Antibodies 101: Affinity Tags

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    Blog Post
    ...This is where affinity tags come in – they are peptides which do not naturally exist in most expression...

  50. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...transduction strategy is to fuse a cell penetrating peptide (CPP) to a protein of interest thereby giving it...
Showing: 1 - 50 of 117 results