We narrowed to 883 results for: prot

...dissecting that S. pyogenes protein into three parts. Their efforts yielded a protein, which they call SpyLigase...for this tool? Howarth: Polyproteins are useful for mechanical study of proteins by atomic force microscopy...introduced the SpyTag/SpyCatcher system, based on a protein isolated from Streptococcus pyogenes that locks...locks itself together, to produce irreversible protein-peptide interactions. In a study published in Proceedings...SpyTag/SpyCatcher system, to be able to grab on to proteins irreversibly in a genetically encoded way. SpyTag...small, just 13 amino acids, but SpyCatcher is a protein. You can fuse SpyCatcher onto things, but even ... advantage of peptide tags for building larger protein assemblies? Howarth: Peptide tags can be very generic...

...extracellular stimuli. One way they do so is through G protein-coupled receptors (GPCRs), receptors that play ...TRUPATH to interrogate 14 of the 16 non-visual G protein signal transducers (collectively known as the transducerome... Gγ), collectively known as a heterotrimeric G protein. This complex converts a signal received by a GPCR... Figure 1: TRUPATH measures heterotrimeric G protein dissociation by bioluminescence resonance energy...the GFP2 tag is fused to the N-terminus of the protein. For the Gα constructs, localization of the Rluc8...challenging. The Roth lab used structure-guided protein engineering and experimental refinement to determine...Rluc8-Gα construct for two of the 16 non-visual G protein transducers. To use the TRUPATH system, cells are...

...CLIP-SNAP fusion proteins for single-molecule FRET. A) Schematic of a CLIP-SNAP fusion protein (a.k.a. CLAP... a flexible linker where a protein of interest can be inserted. B) Protein expression and labeling procedure...optimized strategy uses new combinations of PE protein components (Figure 1), an altered epegRNA delivered...study the conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of...brighter and more photostable than fluorescent proteins, making them ideal for single-molecule fluorescence...FRET to study the intramolecular dynamics of the proteins in solution and when associated with microtubules...generating other SNAPf-, CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids ...

...Non-redundant protein sequences (nr)” database as it contains the largest number of protein sequences. Just...between a Standard Protein BLAST (blastp) and blastx search to verify expected protein sequence in a plasmid... or fusion protein. If your sequencing primer was chosen to confirm a tag or fusion protein is in-frame...of nucleotide or protein sequences. We also use COBALT for aligning multiple protein sequences, particularly...use Standard Nucleotide BLAST (blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). ...or blastx to compare our sequencing results to protein sequences to check open reading frames (ORFs) and...customization. Similar to nucleotide sequences, proteins often have repeated or highly homologous regions...

...endogenous proteins, making the recombinant protein its major secretion. Beyond sheer protein production...expression of recombinant proteins by the pharmaceutical and biotech industries as a protein expression workhorse...is highly efficient at protein secretion, making harvesting recombinant proteins a cinch. This is because...phaffii in the 1970s with the goal of producing high protein animal feed. Due to increasing prices of methanol...of different uses for the phaffii-lous strain. Protein expression   K. phaffii first gained attention ...workhorse. This yeast can also give more protein bang for your media buck - it grows at higher cell densities...post-translational modification of recombinant protein products, specifically glycosylation. The native...

...By tagging proteins with a 7kD degron called mAID and expressing the essential plant protein TIR1 in non-plant...   New toolkit for N or C terminal tagging of proteins for degradation using AID Article contributed ...auxins to conditionally and reversibly degrade proteins of interest fused to a degron, a domain that induces...developed a CRISPR/Cas9 based method to tag endogenous proteins in HCT116 cells with mAID. Using this method, ...generate conditional alleles of a few essential proteins (Natsume et al., 2016). Now, the Kanemaki lab ... plasmids for the N- or C- terminal tagging of proteins with mAID and other tags (Yesbolatova et al., ...CRISPR. This toolkit makes it easy to tag your protein of interest to create conditional human cell lines...

...acid sequence found in the protein, or, are injected with the whole protein. After isolating the B cells...specific region of your protein of interest. For example, if you are studying a protein that may bind a ligand...antibodies against many types of molecules including proteins, lipids, and carbohydrates, in this blog post ...post we’ll focus on antibodies that recognize proteins. Figure 1: Steps in the generation of hybridomas...animals that were immunized against the target protein. Animals are first injected with a small peptide...2016). If your antibody recognizes a part of your protein that may be masked while the ligand is bound, then...negative signal - the antibody can’t recognize the protein even though it is present in your sample. Additionally...

...activator protein cannot bind. After an inducer binds to the activator protein, the activator protein can bind...repressor protein actively prevents transcription. Once an inducer binds the repressor protein, the repressor...repressor protein is removed from the DNA. With the repressor protein absent, transcription is turned ON...used for bacterial protein purification. When arabinose is absent, regulatory protein AraC binds O and ... and largely defines the amount of recombinant protein that will be produced. Many common promoters. like...promoter requires removal of the lac repressor (lacI protein) for transcription to be activated. In the presence... arabinose, cAMP complexed with cAMP activator protein (CAP) can also stimulate AraC binding to I1 and...

...ankyrin repeat proteins, or DARPins, are genetically engineered small affinity proteins, around 14-15 ...derived from a class of binding proteins known as ankyrin repeat proteins. DARPins typically consist of ...the ability to specifically recognize and bind proteins. They’re used in a number of research and clinical...re talking about a standard antibody: An ~150kD protein with both light and heavy chains made by immune... variables (scFvs) are ~27kD engineered fusion proteins similar to a Fab fragment but created through ...scFvs with a CH3 linker, resulting in an ~50kD protein. Minibodies are bispecific, meaning that each scFv...applications, including imaging.  Diabodies are ~60 kDa proteins formed by two Fab fragments bound by short peptide...

...Activity of Protein Variants on a Large Scale Using Deep Mutational Scanning.” Nature Protocols 9.9 (2014... test how multiple amino acid changes affect a protein of interest’s function. This technique relies on...library that expresses all desired variants of a protein. Applying a selective pressure winnows the pool...the ability to generate full libraries of mutant proteins. Researchers from the Whitehead lab developed ... mutational scanning Hundreds of thousands of protein variants can be tested in one experiment. There...possibly discover unexpected epistatic changes in protein activity. See Natarajan et al for an example of...reaction. Below is a step-by-step overview of the protocol: Prepare ssDNA template: The wild-type plasmid...

...Resources on addgene.org Western Blot Protocol Western Blot Protocol Video ...Western blots are a great tool to identify a protein of interest in a complicated solution like cell ...— and what if you want to detect more than one protein in your sample? Or what if something weird happened...membrane, which is sturdier and retains the blotted protein better than a nitrocellulose membrane. If you have...probe for. Even using PVDF, some of your target protein will be removed from the membrane along with the...methods. If you are planning to detect multiple proteins through several rounds of stripping and reprobing...your signal by starting with the least-abundant protein and working toward the most abundant. If the abundance...

...published a protocol that would work for your system. Addgene has depositor submitted protocols and links...transgenic embryos. Cas9-gRNA ribonucleoprotein complexes Purified Cas9 protein and in vitro transcribed gRNA...Single cells Cargo Nucleic acids or protein Nucleic acids or protein Throughput High Very low Efficiency...Do you want to create a fusion with a reporter protein such as GFP? All of these outcomes can be most ...deliver the components as DNA, or would mRNA or protein delivery be more suitable? Determine how you will...nucleofection, viral transduction or injection as either protein, RNA, or DNA. Finally, once you have identified...components, expression decreases as gRNA and Cas9 protein are degraded within the cell.   Delivery methods...

...the viral protein reverse transcriptase (RT). In the case of a natural virus, both the protein and the ...the fact that the virus must have its structural proteins expressed from a reverse-transcribed piece of ...gene therapy vector! For viral vectors, only the protein for reverse transcriptase (RT) is present in the...may seem like a mystery. But it’s simple. The RT protein is made by a separate set of instructions in the...components of the vector particle, some of the RT protein gets packaged inside of the vector particle, along...of the Central Dogma of Biology (gDNA → mRNA → protein): their roadmap includes RNA → DNA (Cooper & Temin...retroviral vectors can still direct the translation of proteins because they integrate this reverse-transcribed...

...identify protein and nucleic acid binding partners at numerous genomic loci through proteomics and the ...al., 2013). However, the fusion of dCas9 with a protein tag allows for the isolation of a genomic region...DNA-binding molecule, such as Zinc-finger or TAL proteins, can be used. The Fujii lab demonstrated this ...spectroscopy studies allowed for the identification of proteins associated with the IRF-1 locus. Figure...that do not use dCas9, require Zinc-finger or TAL proteins targeting a specific genomic locus, which can ... addition to the identification of DNA binding proteins, this method can characterize nucleic acid-nucleic...CRISPR complex, consisting of a recombinant dCas9 protein and synthetic gRNA, is incubated with fragmented...

...prime editing and found the protein La (a small RNA-binding exonuclease protection factor) promotes editing...Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] and Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target distinct...the SARS-CoV-2 nucelocapsid protein, one of the virus’s most abundant proteins and a critical component ...SARS-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M]...transcriptase enzymes, inhibition of host cell proteins, and modifications to the pegRNA. The Britt Adamson...stabilizing the 3’ end of the pegRNA, which is not well protected by Cas9. This is similar to the proposed mechanism...prime editing with an endogenous small RNA-binding protein. Nature. 2024 Apr 3. doi: https://doi.org/10.1038...

...capsid proteins are analyzed. The abundance of viral capsid proteins as a fraction of total protein present...electrophoresis. Viral capsid proteins VP1, VP2, and VP3 are shown relative to the total protein present in the sample...distributed to customers. Details about our production protocols, titering methods, and quality control are described...elements and an internal control of known titer (protocol modified from Lock et al., 2014.) Previous experience...vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC). ...by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic properties of the...relative stoichiometric ratios of the viral capsid proteins VP1, VP2, and VP3. Samples of viral preparations...

...two parts: 1) an activatable protein (the FAP - fluorogen activating protein - dL5** in this case), and ...you can do just that.  Photosensitizing dyes and proteins have been around for awhile (check out this review...recently developed a photosensitizer composed of the protein complex complex and the MG-2I-dL5** fluorogen that...few definitions. A photosensitizer is a dye or protein that absorbs light to produce reactive oxygen species...produced by photosensitizers inactivate target proteins or cells while having little effect on neighboring...fluorogen is not fluorescent until it binds its protein partner thus forming the full FAP-TAP complex (...suggests that it could be used with these fluorescent proteins. He et al also compared MG-2I-dL5** head-to-head...

...viral proteins in human cells and used these viral “bait” proteins to fish for human “prey” proteins that...UCSF). Our approach is to discover key human proteins that can be targeted (hopefully by existing therapeutics...the virus. As a result, we identified 332 human proteins that the virus interacts with in cells. Because...Because viruses rely on the help of human proteins in their lifecycle, identifying these potential human helpers...better, more than half the drugs hitting these proteins are FDA-approved or in clinical trials or preclinical...understanding how SARS-CoV-2 interacts with human proteins. For example, we used HEK293T cells in our first...set of constructs comprises all SARS-CoV-2 viral proteins except for Nsp3 and Nsp16, and includes an additional...

...streak plate challenge. Chromoproteins are a subset of the fluorescent protein family that give off color...? Find the chromoprotein expressing plasmids here, and check out our streak plate protocol video on Youtube...Addgene.org Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids at Addgene Check out...outcompeted the other species.  When he found the chromoprotein plasmids available from Addgene, he knew it ...color in ambient light. Expressing these chromoproteins in E. coli would make it easy to distinguish colonies...addition to streaking for single colonies, these chromoprotein-expressing strains are easily amenable to other...blog Read our other blog posts on fluorescent proteins Check out educational resources on the blog Resources...

...GFP-nanobody rather than a specific protein of interest. This fusion protein can then be co-expressed in essentially...stabilizes the TurboID-nanobody protein and allows TurboID to start labeling proteins in close proximity to the...Proximity-dependent labeling has become a powerful approach for proteomic mapping in both cell culture and whole organisms...approach has been hampered by the need to tag each protein of interest and, for in vivo work, generate a dedicated...zebrafish line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to...the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The modularity of...including limitations of number of flourescent proteins that can be inserted into a cell without disrupting...