We narrowed to 883 results for: prot

...distinguish between proteins bound by the antibody vs proteins bound to the target protein, so it’s important...your antibody is not binding to proteins outside of your target protein. Show suitability for an intended...of a protein of interest. Conveniently, you found a cell line in which the gene for your protein has been...express your protein of interest endogenously and transfect them with a plasmid encoding your protein of interest...expected target protein in a Western blot. You acquire a plasmid encoding your protein of interest, known... the majority of the protein in the sample was identified as your target protein, allowing you to proceed...methods may include analysis of transcriptomic or proteomic data from several different tissues or cell types...

... Abundant (LEA) proteins, small proteins that naturally adsorb to the AWI, can protect cryo-EM samples...enrichment and structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered... of proteins captured on magnetic beads (Arimura et al., 2024). In MagIC-cryo-EM, target proteins (expressed...doi.org/10.1101/2024.01.21.576499.    LEA proteins protect fragile samples during cryo-EM grid plunge...complexes may dissociate  (right). B) Protein samples protected from AWI by helical LEAs. C-E) cryo-EM.... J. (2024). Small LEA proteins as an effective air-water interface protectant for fragile samples during... DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription...

...Biotin protein ligase (BPL) is responsible for this highly specific post-translational protein modification...the lab are protein purification and immunoprecipitation experiments. Biotinylated proteins or complexes...terminus of the protein. BirA has been shown to effectively biotinylate AviTag expressing proteins in bacteria...fusion proteins Learn how biotin is used in Split-BioID: An improved method for studying protein-protein...biology for an array of different techniques and protocols. In this post we will discuss the natural role...about in vivo biotinylation of bacterial fusion proteins Biotin is an essential vitamin for life Although... general process by which biotin is added to a protein or macromolecule. Biotin can be added enzymatically...

... Which Fluorescent Protein Should I Use? However, fusion proteins tag your protein of interest with a ...target protein composition. It is used for a variety of applications such as measuring target protein expression...produces a greater SSC.  Protein expression, measured using fluorescent protein fusions In addition to ...-stained protein will emit a stronger photon than a cell expressing low levels of the protein. The electrical...on the N- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the ...structure of the protein. In some cases this can lead to changes or a complete loss of protein function. In...the challenges of fluorescent protein fusions by binding to the protein in its native state.  Antibodies-based...

...amplification protocol developed for each individual pooled library; generalized amplification protocols tend ...reference protocols There are many decisions to make when developing your amplification protocol! We recommend...one-size-fits-all” protocol! Each library is different, and you should always experimentally test the protocol you design...surprised to learn that we ask for an amplification protocol with your deposit. This is because repeated amplifications... we’ll discuss how to develop an amplification protocol for your pooled library deposit. When a requesting...consider when creating a pooled library amplification protocol. Created with BioRender.com.   The most important...efficiency you need to achieve. Most pooled library protocols aim for somewhere between 30x and 200x coverage...

...)est fluorescent protein Fluorescent Proteins 101: A History of Fluorescent Proteins ...attached to self-labeling proteins. Some common examples of these self-labeling proteins are SNAP-tag® and HaloTag...to using these self-labeling proteins over traditional fluorescent protein tags, such as high specificity...many self-labeling protein fusions as well as empty backbones to clone your own protein of interest. Fluorescence.... FITC binds to an amine on a protein, enabling the labeling of protein substrates/peptides and antibodies... as conjugation with proteins, should be carefully managed. Proper dye-to-protein ratios, or the degree...cleavage of FITC-conjugated protein ligands is unaffected by additional protein from cells under certain ...

...insight into protein:DNA interactions. ChIP works by using antibodies to capture protein:DNA complexes with...to your protein(s) of interest. Once the complex is captured, it is treated so that the protein and DNA... antibody on crosslinked proteins. In the case of histone modifying proteins, which bind quite strongly...with RNAse and Proteinase K to purify the DNA and then determine concentration.  Protein Capture After ...that ChIP uses native proteins (so antibodies validated only for denatured proteins may not be a good choice...specifically select for proteins of interest (4) isolated your antibody:protein:DNA complexes and finally... to firmly attach the proteins to the DNA, depending on how tightly the proteins bind natively. But the...

...permeabilization protocol, but very poorly in a methanol fixation-permeabilization protocol. Although the... the optimal knockdown of the protein target will depend on the protein turnover rate. If you need a more...suppress the expression of many related proteins, not just your protein of interest, which can affect your ... levels of the target protein. It may only be able to detect the target protein at the high levels caused...Additionally, their performance depends on the specific protocol used and the abundance of the epitope in the sample...can detect the antigen of interest in the given protocol. This requires use of one of the five pillars ... of which are commercially available. Various proteomic and transcriptomic datasets can be used to select...

...C terminus of a protein may disrupt the biological function of some proteins. Protein folding issues, ...of detection of a mutant protein vs. endogenous WT levels of the same protein when both are expressed ...to a protein’s termini to increase detection. The tag can also be removed by treating the protein with...validate tagged protein functionality (Arribere, et al). In rare cases, tags can also affect protein localization...for your application, specifically detects your protein, is species compatible, and doesn’t come with a...peptides introduced at the N or C terminus of a protein that are bound by antibodies for the purpose of...of purification or detection of the tagged protein. These peptides consist of as little as 8 or as many...

...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...