We narrowed to 695 results for: prot

...producing both a tagged protein from the tagged allele and an untagged protein from the untagged allele...of the protein gleaned from the literature and engagement with researchers who study the protein or structure...possible. This is especially true in the case of protein tagging to elucidate cellular structures. Overexpression...Overexpression methods flood the cell with protein, which can both interfere with a cell’s normal function... hard to visualize the precise location of the protein or structure of interest. Endogenous gene tagging...tagging and visualization of specific, individual proteins under endogenous regulatory control. But even ...transcribed and translated. For an N-terminally tagged protein, we used the same strategy, inserting the tag and...

...encodes a single envelope protein called rabies glycoprotein (G). G-protein-coated RABV infects cells ...order neurons, via entry mediated by the G protein/G-protein receptor. Once in first order neurons, the...recombinant rabies virus vectors. Nat Protoc. 2013;8:1583–1601. doi: 10.1038/nprot.2013.094. Pubmed PMID 23887178...line. By doing this, RABV expressing the viral protein EnvA on its surface will only infect Cre-positive...into these phenomenon has shown that the RABV G protein is necessary and sufficient for retrograde transmission...can be controlled by selectively supplying the G protein in trans only to starter cells (infected with G-deleted...cell. RABV replication will then occur and the G protein (supplied in trans) will be incorporated into the...

...GAG-POL polyprotein Nanoblades are formed when viral structural proteins, such as the MLV protein Gag, multimerize...the MLV Gag-Pol polyprotein, cleaves the proteolytic site connecting the two proteins, thus releasing ...purified by centrifugation. Gag-Cas9 fusion protein The MLV Gag protein fused to a Flag-tagged SpCas9 is incorporated...the unmodified MLV Gag protein. After Gag-Cas9 is incorporated, the MLV protease, supplied as part of the... envelope proteins To alter the cell tropism of Nanoblades, different viral envelope protein(s) are expressed...MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named...with the the Cas9 portion of the Gag-Cas9 fusion protein. A single Nanoblade particle can be loaded with...

...milk protocol is adapted from the 1979 Patterson Protocol. When published, the Patterson Protocol was ...causes the proteins and genomic DNA to precipitate. Plasmid DNA is free in solution. Proteins and genomic...plasmid purification protocol and guanidine isothiocyanate for the gel extraction protocol) disrupts the DNA...the same principles: cells are lysed, RNAs and proteins are removed to leave you with DNA. This DNA is...Product: plasmid DNA This kit-free plasmid miniprep protocol from Addgene follows a similar workflow as a column-based...you lyse the bacteria and denature the DNA and proteins in solution. The pH is then lowered using a renaturing...phenol chloroform extraction to remove remaining proteins and RNase. Finally, DNA is precipitated with alcohol...

... are trying to remove from your sample is protein. Proteins absorb light to varying degrees across the...specific to proteins and why researchers traditionally use the A260/280 ratio as an indicator of protein contamination...DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol    ...downstream experiments. The usual suspects, such as proteins, phenol, or guanidine salts, exhibit their own...how different contaminants impact these ratios.  Protein contaminants Whether you are purifying plasmid ... The A260/230 ratio can be a good indicator of protein contamination, but a number of other potential ...A260/280 ratio does not drop significantly until protein levels exceed 70% of the sample content (Loughrey...

...on designing entirely new proteins, improving our understanding of how protein folding works and creating... publish a human-designed protein that was not based on any existing protein structures, called Top7. ...novo designed proteins also helped researchers understand more about the biophysics of protein folding and....com. Top7 gets its name from “topology of protein” diagrams, which show the interactions between ...closer to achieving it. Once they designed the protein computationally, the researchers expressed, purified... a powerful proof of concept that this kind of protein design is possible. Although Top7 was designed ...created it have been applied to numerous other protein design goals, both from the Baker Lab and others...

... and a protein standard. The density of the antibody bands can be measured against the protein standard... Antibody Purification Protocol Addgene's Video Antibody Purification Protocol Addgene's Antibody Guide...predict the molar absorption coefficient of a protein. Protein Sci. 1995 Nov;4(11):2411-23. doi: 10.1002/...applications you are planning, cost, and the amount of protein you need to produce.  The most common yeast strain...pastoris) because it secretes fewer of its own proteins, making the purification process simpler. Due ...incubation time to maintain cell viability and increase protein production (Schwarz et. al., 2020). Harvest and...quantification. Many spectrophotometers have an IgG protein setting. If yours does not, be sure to set the ...

...decorated with a fluorescent protein remain functional. Attaching the fluorescent protein fusions allowed us to...produced by orchestrated movements of networks of proteins consuming molecular fuel sources. While the importance... can be generated by conformational changes in proteins? How can information propagate through a structured...rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony explanation...addressed using R bodies: simple, force-generating protein machines that can extend and retract over many ...

...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs... including popular new approaches like PROTAC targeted protein degraders. But CRBN has been difficult ... produced by our partners at the Institute for Protein Innovation (IPI), part of a new collection of epitope...short, linear epitope derived from the P and V proteins of Simian virus 5 (SV5) (Randall et al., 1987)...drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as a promising tool for...future structure-guided drug design and studies of protein ubiquitination and degradation.   Find the CRBNmidi...for crystallographic and biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org...

...include proteins like nonstructural proteins 1 to 16, spike proteins, envelope small proteins, and membrane... Currently, plasmids containing all SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression...mammalian expression vectors containing various spike protein variants are also available. Plasmids are continually...membrane proteins. Although all viral components are available in bacterial expression plasmids, glycoproteins...glycoproteins and other proteins that require post-translational modification may not be easily expressed ...plasmids The yeast expression constructs contain protein coding genes, tags, truncations, and codon optimizations...level expression and purification of SARS-CoV-2 proteins for downstream experiments and offer flexibility...

...specifically curated for protein expression in K. phaffii with special attention to protein modifications which...successfully for protein localization and transport studies in HEK cells, recombinant protein purification...great option for those interested in recombinant protein purification and metabolic engineering. If instead...library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent option (Iverson...compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes...eukaryotes! The technology has been used to optimize protein secretion and expression in the biotech workhorse...formerly known as Pichia pastoris). If your goal is protein isolation, then the MoClo Pichia Kit will get you...

...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...

...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...

...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...

...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...