We narrowed to 883 results for: prot

...specific protocol for producing virus, I recommend that you follow that protocol. If not, our protocol on Lentivirus...the protocols provided by the depositing lab whenever available. Addgene also shares the protocols we’...these are not general amplification protocols. Rather, each protocol is optimized for a single, specific...amplification protocol, or have constructed your own library, you’ll need to develop your own protocol that amplifies...lentiviral CRISPR libraries. Protocols from the Broad Institute- A variety of protocols and guides regarding ...activation screening. Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. Epub 2017  ...with a colony formation titering assay or similar protocol. After functional titer is determined, larger ...

... in both E. coli and S. cerevisiae. Protein folding - If a protein is encoded by a mixture of codons with...resulting in the production of nonfunctional proteins and protein fragments respectively. Solving the problem...every region of a protein should necessarily be translated rapidly to produce a protein that functions properly... rate of full‐length human proteins expressed in Escherichia coli." Protein Science 19.7 (2010): 1312-... blog Plasmids 101: protein expression Plasmids 101: E. coli strains for protein expression Plasmids ...Controlling gene expression through gene sequence - Proteins that are encoded by codons with low abundance ...charged tRNAs may be produced at a lower rate than proteins encoded by highly abundant, charged tRNAs. For...

...About Other Protein Tags Catch Up on Your Fluorescent Protein Background Which Fluorescent Protein Should ...Fluorescent Protein Fusions Request Plasmids from the Michael Davidson Collection Find Fluorescent Protein Kits... chemistry! The discovery of green fluorescent protein (GFP) sparked a renaissance in biological imaging... their own. Subsequent advances in fluorescent proteins have replicated many of the properties once exclusive...example, moving from transient overexpression of protein fusions to gene-edited cells can decrease the number...budgets are a widespread issue with fluorescent proteins—even the thriftiest cell biologist can feel like...brighter and more photostable than fluorescent proteins, providing a straightforward way to improve the...

...AID-tagged protein. To quantify protein degradation the Ward lab used fluorescent proteins (FPs) fused...of Golden Gate cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows ...implications for the fusion protein effectivity), and a fluorescent protein tag for visualization (Fig....chromatin-modifying proteins quickly and accurately.   Priode and Haynes, protocols.io. 2021. dx.doi.org...attached to your protein of interest. TIR1 interacts with endogenous Skp1 and Cul3 proteins to form a SCF...inducible degron sequence) tagged proteins. Degradation of your protein interest would also result in depletion... histone-modifying enzymes and reader-effector proteins. b) Golden Gate assembly (backbones available ...

...only be added to the C-terminus of proteins of interest and proteins tagged with it needed to be chemically...1986) An hsp70-like protein in the ER: Identity with the 78 kd glucose-regulated protein and immunoglobulin...commonly used Myc-tag. Who first started using it in protein tagging experiments? Why Myc? When did the commonly...the life and times of the myc tag.  Origins of protein tags For about as long as the disciplines have ...antibodies as tools allowed us to detect and capture proteins of interest to better study them in vivo and in...they landed on was to recombinantly append their proteins of interest with unrelated peptides that already...peptide from the neuropeptide substance P. This protein had a highly specific antibody to the C-terminal...

...the name "G-protein coupled receptor" into two parts: the receptor itself and the G-proteins it associates...Takeda et al., 2002). G proteins: The receptor is coupled with heterotrimeric G proteins on the intracellular...beta-arrestin-TEV protease fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription... of the heterotrimeric G protein complex that can be used to detect G protein activation via bioluminescence... catalyzes the biotinylation of proteins near the GPCR. These proteins are then identified with mass spec...besides being one of the five senses? GPCRs or G-protein-coupled receptors (GPCRs)! Aside from these three...recognizes the appropriate ligand to activate the G proteins. It is estimated that there are approximately ...

...Proximity labeling can identify protein interactions or map a local proteome, but some applications have ...locations. C) Enrichment of dual-labeled proteins identifies proteins that had been trafficked from the “source...generated a fusion protein called iPAK4 that assembles into a slow-growing protein fiber in the cytoplasm... Figure 4: Time-resolved protein ticker tape. A) Design of iPAK4 protein ticker tape recording. B) HEK...labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old antibody new tricks...recently used phage-assisted continuous evolution and protein engineering to generate seven new prime editors...., et al. (2023). Phage-assisted evolution and protein engineering yield compact, efficient prime editors...

...efficiency of tobacco etch virus protease (TEV), a highly sequence-specific protease used to cleave affinity tags...surface) which enables tracking of protease activity with a fluorescent protein--no affinity purification required...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor...'s what you'll find in this post: Yeast-based protein evolution Fluorescent tools for mammalian cells...viral service Faster catalysis after yeast-based protein evolution By Joanne Kamens Mateo Sanchez and Alice...sequences that evolve a measurable property of a protein. As a proof of concept, Sanchez and Ting sought... tags from purified recombinant proteins at small and large scale.  This new technique involves selection...

...Indirect labeling is achieved through protein-tag or protein-protein interactions, like biotin-streptavidin...transmembrane proteins, such as receptors, or post-translational modifications on those proteins, like glycosylation...kits allow for labeling of cytosolic proteins, as well as proteins of the secretory pathway. Those can ...buffer to label intranuclear proteins of interest, you can label cytosolic proteins alongside the intranuclear...like many of the cluster of differentiation (CD) proteins found on immune cells. However, there are more...methods can include labeling markers with multiple proteins/antibodies, targeting markers found within the...cytosol/nucleus, or visualizing DNA instead of proteins. Some stains can diffuse through the cell membrane...

...fluorescent protein earlier described. “Until our proteins, there were lots of fluorescent proteins made from...you start poking around on Addgene’s Fluorescent Protein Guide to In Vivo Imaging, you’ll pretty quickly.... We all know scientists have used fluorescent proteins to observe what’s happening inside cells for at...decades. Green is the classic color, but fluorescent proteins are available in a variety of hues. While those... The key to their bacterial phytochome-derived proteins, iRFP670, iRFP682, iRFP702, iRFP713 and iRFP720... The challenge then was to develop fluorescent proteins that would fall within that near-infrared range... and that’s exactly what iRFPs achieved. Those proteins allowed a signal-to-background ratio in mammalian...

...surface proteins with a membrane-impermeable biotin substrate (Figure 1). Biotinylated proteins were isolated...In addition to unique proteins on each cell type, the cell-surface shared proteome of astrocytes and neurons...genetically fused to a protein of interest or targeted to a GFP fusion protein using an anti-GFP nanobody...and translation of chromoprotein and gRNA plasmids. Nuclease-dead Cas9 (dCas9) protein is supplied separately...Here's what you'll find in this post: Cell-surface protein labeling tools for neurons and astrocytes Nerve-to-cancer...Frugal CRISPR kit for classrooms Cell-surface proteomics for neurons and astrocytes By Mike Lacy Cell-...Khakh and colleagues mapped their cell-surface proteomes (Wu et al., 2025). The team expressed extracellularly-anchored...

...recruits host proteins to the ori to initiate replication. At high copy numbers, RepA proteins form complexes...when they depend on the host cell’s initiation proteins to start their own replication. In general, these... begins. This process relies on the initiation protein DnaA. First, DnaA must bind ATP. Then, the ATP-...bound, including at the low-affinity sites, the proteins work cooperatively to separate nearby AT-rich ... available DnaA box. In bacteria, however, the protein integration host factor (IHF) assists with the ...DnaA boxes are occupied by ATP-bound DnaA, the proteins cooperatively can separate the DNA strands. Created...does get hydrolyzed over time, inactivating the protein binding it (Grimwade et al., 2018). Rather than...

...major capsid protein VP1 is co-expressed with a fusion protein containing VP2C and a protein of interest...jbc.RA120.01289     Novel protein cage provides simple enzyme protection in yeast bioproduction pathways...MPyV VLPs ). This artificial nanoparticle can protect proteins from unfavorable interactions in the cell,...fusion protein anchors to VP1, which self-assembles into a compartment encapsulating the protein of interest...receptor binding domain (RBD) of the viral spike protein.   Figure 1. (A) SARS-CoV-2 RBD DMS Library...enzyme 2 (ACE2) cell-surface receptor. The spike protein RBD responsible for binding ACE2 is the target ...library, each plasmid encodes the mutated spike protein and a barcode, and is designed so that the SARS-CoV...

... localization of individual proteins, but also investigate protein-protein interactions via FRET. Sidae... to integrate protein tags into the genomes of S. cerevisiae and S. pombe. The protocol is surprisingly...epitope tags, protease sites, and selection markers. These are especially useful in protein complex studies...combination of protease site (TEV or PreScission), epitope tag (12xHis, 2xStrep, 3xFlag, Protein A, or V5),... unique protein fusions. This collection is ideal for tandem affinity purification of protein complexes.... A given insert will typically contain both a protein modification sequence and a selectable gene product...studies where tagging of multiple protein products is desired, as multiple selection markers can ensure ...

... domain structure. Glypicans are proteoglycans composed of a protein core, 2–4 glycosaminoglycans (GAG...mini counterpart label nearby proteins with biotin. The biotinylated proteins can then be isolated and identified...for Glypicans DNA editing with STITCHR Dynamic protein labeling with TurboCas Enhancer-AAV viral preps...after inserting EGFP and provide step-by-step protocols for viral generation and stable cell line creation...our bacterial plasmid set for expressing tagged proteins, and stay tuned for mammalian expression and AAV...therapeutic potential. Our partners at the Institute for Protein Innovation (IPI) recently released a set of highly-specific...antibodies against three of the six human glypican proteins (GPC1, GPC3, and GPC4). All IPI antibodies undergo...

...subjected to the experimental protocol but does not receive either protein kinase inhibitor. Since the ...example, if you are studying the effect of a new tau protein kinase inhibitor on slowing diseases progression... you will be able to confirm that the testing protocols elicited the expected response and have a baseline...experimental (?) groups were run on a western against a protein standard (P) to check for tau molecular weight ...included to assess variability in the experimental protocol. In the experiment above, you could repeat the...knockout samples are treated with the identical protocols that the experimental samples undergo. If the ...repeated for each experimental run as long as the protocol remains the same.    Figure 2: Microscopy-based...

...genetically encoded peptide (DogTag) and protein (DogCatcher); ideal for protein loop regions CRISPaint Gene Tagging...HUH-tag For creating DNA-protein conjugates rHUH HUH-tag (RNA) Covalent protein tag for RNA labeling ...University of Cambridge. His research focuses on protein innovations for therapeutic and vaccine design....be similarly useful across the diverse kinds of proteins that one finds in cells. High yield of reaction...development of genetically engineered components, like protein tags (Figure 2). Through the work of many groups...inteins or SpyTag, which are useful for connecting proteins in the test tube or in cells. There are also SNAP-tag...SNAP-tag and HaloTag modules, which can connect proteins to any kind of small molecule probes with different...

...labeling excitatory inhibitory synaptic proteins Fluorescent protein system for C. elegans Fluorescent timers...fluorescent proteins system for Caenorhabditis elegans By Jennifer Tsang Fluorescent proteins (FPs) traditionally...the target protein. The team validated this tool by targeting six different genes whose protein products... Labeling excitatory and inhibitory synaptic proteins with FingRs By Jason Nasse Since the pioneering...neurotransmitters. Using genetically encoded fluorescent proteins coupled with Fibronectin intrabodies generated...FingR constructs target the synaptic scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory...neuron, the FingRs bind to the endogenous synaptic proteins allowing them to localize to the synapse and minimize...

...components: a GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP...NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP... PP7 and MS2 coat protein binding sites to tether many copies of the FP-coat protein fusions to their ...RFP-labeled osk and Oskar protein were detected at the posterior pole. Oskar protein levels were negatively...Brightest Fluorescent Protein Resources on Addgene.org Find Fluorescent Protein Plasmids Check Out Other...mRNAs are digested with ribonucleases, ribosomes protect the segments of mRNA to which they are bound. These... to monitor local translation and determine if proteins are targeted to their destination co- or post-...

...about Photoactivatable Fluorescent Proteins Check Out Fluorescent Protein Timers Use Rosella to Study Autophagy...Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein Tags  ...be limited by a few caveats. Since fluorescent proteins (FP) require external light activation, you can...practical due to the low intensity of luminescent proteins. To make luminescent imaging a reality, Addgene...including optogenetics, biosensors, and fusion proteins. Solving the problems of low luminescence By...generates much less signal than comparable fluorescent proteins. This weak emission hasn’t stopped some scientists...scientists from using chemiluminescent proteins for imaging, but it has precluded widespread adoption of the...