We narrowed to 695 results for: prot

...selective removal of proteins-of-interest from the proteome. Rather than inhibiting protein function, these...with the target protein, leading to polyubiquitination of the target protein and proteasomal degradation....molecule degraders” recruit the proteasome to destroy targeted proteins. We previously developed small ...endogenous proteins is an exciting strategy, it is difficult to apply this technique to oncoproteins that lack...co-opt the endogenous ubiquitin proteasome machinery to degrade any protein. We are particularly excited ...fellow at Dana-Farber Cancer Institute. Targeted protein degradation In the Bradner and Gray labs, we synthesize...that achieve selective degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and...

...CRY2) and calcium and integrin-binding protein 1 (CIB1) proteins. The goal of both groups was to create...composed of two fusion proteins: 1) the genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1...form a functional protein due to being fused with high-affinity interacting proteins Coh2 and DocS, respectively...chimeric protein consisting of Cas9 and a photo-dissociable dimeric green fluorescent protein (pdDronpa... saCas9 protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly...light-removable protecting group, specifically a nitrobenzyl photocaged lysine (PCK), on the Cas9 protein (Hemphill...The use of nickases, prime editing, anti-CRISPR proteins, and other techniques all aim to improve targeting...

...visualizing protein turnover Download the fluorescent protein eBook Browse our fluorescent protein blog posts...powerful tool to study proteins in vivo, it does have some limitations. GFP has long protein maturation times... nascent proteins. In addition GFP tags are large, dim, and permanently attached to a protein, making ...different protein sequences called epitopes and scientists can use these to tag their proteins of interest...visualize proteins of interest tagged with the HA epitope, the lab fused a fluorescent protein to the hybrid... of the fluorescent protein allows scientists to easily observe the tagged protein upon probe binding ...also bind a 1xHA tagged histone protein as well as a mitochondrial protein tagged with spaghetti monster...

...transduced using only protein. Read on for a description of the lab’s iTOP protein-only transduction method...common protein-only transduction strategy is to fuse a cell penetrating peptide (CPP) to a protein of interest...in the buffer used for protein purification must be helping the untagged protein enter cells. Subsequent...to determine how much protein iTOP causes a cell to take up, and continued protein expression would require... for off-target effects. Third, protein purification (once a protocol has been established) may be less...Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery Techniques: Introduction...applications to CRISPR/Cas9 genome editing.   Protein-only transduction? Niels Geijsen’s group in Utrecht...

...their fusion to a POI can affect protein folding and protein-protein interactions. Also, overexpression...have created antigenic protein tags called “spaghetti monster” fluorescent proteins (smFPs). smFPs are literally... by red fluorescent proteins such as tdTomato or RFP or blue fluorescent proteins like Cyan. The issue...expressed proteins than standard tags and can be used to distinguish highly similar proteins. For example... W. Piston. "Fluorescent protein tracking and detection: fluorescent protein structure and color variants...Neuronal tracers such as epitope tags and fluorescent proteins (FP) have also been important in mapping, monitoring...short antigenic peptide sequences, attached to a protein of interest (POI), that facilitate immunohistochemistry...

...researchers, from selective protein ligation through introducing ncAAs with protein modifications of interest...’ve shortlisted several protocols, including their phosphoserine ncAA protocol, to optimize and share,... posts on the various protocols and helping to spread the word as new protocols come out! References ...into Recombinant Proteins in Escherichia coli. 12(21). https://doi.org/10.21769/BioProtoc.4541  ...If you study proteins, you’re probably quite interested in the canonical 20 amino acids. But in your ...your quest to learn more about your protein of interest, you may find the available amino acids a bit…limiting... called unnatural amino acids) are placed in a protein site-specifically to alter its function, mimic ...

...experiments. Fluorescent protein fusions One of the easiest ways to track a given protein is to fuse it to a... a fluorescent protein. You can then study where the protein is localized, and how its localization and...overexpress your protein at higher levels, you risk observing artifacts only present when the protein is at a ...Instead of directly fusing a fluorescent protein to your protein of interest, you instead fuse it to the...SunTag’s suitability in your experiments on a protein-by-protein basis. However, since many of Addgene’s SunTag... Up to Receive the Fluorescent Protein eBook Avoid Fluorescent Protein Oligomerization Learn about Small...Addgene.org Find Fluorescent Protein Empy Backbones Visit Our Fluorescent Protein Guide Page Find Luciferase...

...the folded protein (green region). Methanol dehydrates the protein, which can disrupt protein structure...IF, look for general IF protocols for your sample type. Starting with a protocol that usually works in ...details to help you plan and optimize your own protocols. Here we’ll start with just the first couple of... (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction that covalently...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative... polymerize and lose its ability to cross-link proteins. As such, you will get the best results if you...solvents can be quite harsh - lipids and soluble proteins can be lost during fixation with these chemicals...

...effectively separated from DNA and proteins by a simple extraction protocol with guanidinium thiocyanate-phenol-chloroform...Sacchi, 2006).  View our protocol for RNA extraction! Making the simple protocol even more foolproof (still...twenty-something years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot.2006.83 Additional resources...blog Read blog posts related to plasmid protocols Find protocols for purifying DNA without a kit Resources...Addgene.org Check out our molecular biology protocols Find protocol videos on our website Read our Molecular...enzymes that degrade RNA, are especially hardy proteins that are found in and on everything, including...   RNA extraction methods evolved into a simple protocol still used today There are many alternative methods...

.... Spike protein, variants, and pseudotyping The SARS-CoV-2 virus depends on its spike protein to bind ...published a protocol and reagents for pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited...spike protein was developed based on 100 structure-guided spike designs and has improved protein yields...and identified interactions of these proteins with human proteins. Their deposited plasmids expressed ...2020) Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization... more about mapping the SARS-CoV-2 - human protein-protein interaction Find tips for remote collaborations...over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image from Maya Kostman ...

...red fluorescent protein (mCyRFP1). Derived from cyan-excitable organge fluorescent protein (CyOFP1), some...the Doyon lab has also provided a detailed protocol at Protocol Exchange. Plasmid maps can be found at Addgene...specificity, they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish...This New AAV Tool    mCyRFP1: Novel fluorescent protein that enables imaging of two biosensors simultaneously...Podcast Segment New bright monomeric red fluorescent protein - mScarlet! The Dorus Gadella lab has recently ...recently developed a synthetic red fluorescent protein variant they call mScarlet. Bindels et al used a combination...performance in cellular functional imaging and in protein fusions. The Gadella lab has deposited all mScarlet...

...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...