We narrowed to 883 results for: prot

...FRET probes are typically used for studying protein-protein interactions by measuring the change in FRET...acceptor proteins is not optimal, FRET may not occur or be detected even if the two proteins form a complex...processes. Currently, FRET experiments can probe protein-protein interactions, measure the concentration or ...Fluorescent Protein Biosensors Browse the Entire Fluorescent Protein Collection ...Imagine being able to determine whether two proteins are within 10 nanometers of each other, or measure...structure of spider silk, or the activity of a protein in a synapse. What kinds of tools enable us to ...with fluorescent molecules, including fluorescent proteins, has led to the alternative acronym Fluorescence...

... fluorescent HPLC (proteins or other molecules) Isolate or quantify cells, proteins or other molecules...given molecule. The sensor component is usually a protein that undergoes a conformational change in response.... Some biosensors contain a single fluorescent protein fused to a domain that senses the concentration...created by Roger Tsien’s lab contain two fluorescent proteins (a FRET pair) separated by CaM and M13. When calcium...increases FRET transfer between the fluorescent proteins. FRET is highly amenable to biosensor creation...of biosensors To focus on a particular gene or protein, check out our gene-specific biosensors. These ...also use biosensors to follow processes such as protein or DNA binding, enzyme or transcriptional activation...

...oligonucleotides over proteins is their modularity, which is lost as soon as the aptamer is bound to a protein fluor.... of protein-based fluorophores, including: Environments with higher temperatures, since proteins can ...flexibility is their broad range of targets, including proteins, peptides, amino acids, drugs, metal ions, and...incorporate themselves into a larger target, such as a protein. Such flexibility makes aptamers ideal candidates... can be used as powerful biosensors, the bound protein fluorophores and quenchers are still relatively...fluor. Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether... compound malachite green (MG) to create a non-protein based visualization technique. Ultimately, MG proved...

...of a fusion protein is the efficient purification of an already immobilized target protein. The disadvantage...immobilization for kinetic analysis Proteolytic removal of the fusion protein could provide a more relevant ...target protein with the vitamin biotin. The interaction between biotin and the egg-white protein avidin...tetramers of biotinylated protein. However, oligomerization of biotinylated proteins on avidin can be controlled...produces biotin ligase and a protein of interest fused to maltose binding protein (MBP, New England BioLabs...to express a foreign protein in bacteria and subsequently purify the fusion protein from a bacterial extract...can be removed using Xa protease. The carboxyl-terminus of the fusion protein includes an AviTag and a...

... to directly deliver a ribonucleoprotein (RNP), consisting of the Cas9 protein in complex with a targeting...stability of the Cas9 protein in the host. Indeed, research has shown that a Cas9 protein from Geobacillus ...Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and...delivery of, at minimum, two components: the Cas9 protein and a guide RNA (gRNA) targeting your genomic site...translation delays editing What are Cas9-gRNA ribonucleoproteins? One alternative approach, which avoids many...Using RNPs can also alleviate difficulties with protein expression that occur in cells where common eukaryotic...transfection, and are quickly cleared from the cell via protein degradation pathways. This makes Cas9 RNPs useful...

... Addgene Blog Study Protein-DNA Interactions with CUT&RUN Study Protein-Protein Interactions with Split-BioID...like DNA and protein get all the attention.There are numerous ways to detect DNA-protein interactions ...CHIP-seq, FAIRE-seq, Cut & Run) and to detect protein-protein interactions (yeast-two hybrid, Co-IP, BioID...you want to study RNA-protein interactions? The characterization of RNA-protein interactions has lagged...non-physiological RNA-protein interactions due to sample processing. Require prior knowledge that the protein of interest...immunoprecipitation (CLIP) identify RNA-protein interactions by cross-linking RNAs and proteins with UV light, followed...antibody pulldown of a protein of interest, much like using ChIP for studying protein and DNA interactions...

...on the activity of a given protein? Classical methods for studying protein phosphorylation A number of...phosphorylated intermediate of a protein for characterization. However, phosphoproteins are often low abundance...phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular toolkit...To begin, one should optimize protein production using standard protein purification strategies; expression...function, protein interaction, etc. by comparison to non-phosphorylated or ‘wild type’ control protein. The... type protein from the same plasmid.  Upon verifying that serine phosphorylation affects protein function...pSer-incorporated protein, ideally bringing the phenotype assayed closer to that of the wild type protein. The efficiency...

...the proteins they label. In fact, a recent study of 400 proteins found that 20% of tagged proteins did... they compromised protein-protein interactions. If you’re looking to a tag a protein, inntags IT5 and ... native protein structure, inntags may become the tags of choice for examining protein-protein interactions...tag may interfere with protein localization and/or function, so each tagged protein must be tested carefully...Immunofluorescence and fluorescent-protein tagging show high correlation for protein localization in mammalian ... other Protein Tags in our Plasmids 101 Series. Get the Inntag Plasmids Tag endogenous proteins with CRISPR...First described in the 1980s, protein tags are now one of the most useful items in a scientist’s toolbox...

...of integral-membrane protein constructs) protein tag. MISTIC drags tagged proteins into the membrane and... Protein purification can be one of the most stressful lab activities. Working with proteins requires...strains designed for protein expression. Cultures are then lysed and the protein of interest is purified... improve protein stability and solubility. Determining the best way to express one’s protein of interest...bonds in your protein - two parameters that affect how well E. coli can produce your protein. Structural...Structural information for many proteins is easily accessible in the RCSB Protein Data Bank and PDBsum. After ...MBP improves protein solubility and stability. TRX and signal peptide tags help proteins with disulfide...

... photoconvertible proteins are PA-GFP, Dendra2, and the mEOS proteins. Some proteins are reversibly switchable...you really know how fluorescent proteins glow?   Fluorescent Proteins (FPs) were first discovered over...the discovery of the Green Fluorescent Protein (GFP), a protein from the jellyfish Aequorea Victoria. ...categories of fluorescent proteins Photoactivatable / Photoconvertible: These proteins can switch their color... less).  For LSS proteins, the Stokes shift is ≥ 100nm. Specifically, these proteins are excited by UV...when labeling proteins that interact. Indeed, FRET is often used to determine if two proteins interact. ...processes” Curr. Protein. Pept. Sci. 9(4):338. Chudakov et. al. (2010) “Fluorescent proteins and their applications...

...magnetoresponsive proteins. Motivated by curiosity Although a wide range of fluorescent proteins and optogenetic...fluorescent proteins respond to magnetic fields?” and “What are magnetoresponsive fluorescent proteins good ...do just that — by developing a new fluorescent protein tool that responds strongly to magnetic fields ...possibilities of a magnetoresponsive fluorescent protein. Controlling an optogenetic or fluorescent tool...and the interactions of organic molecules like proteins with magnetic fields generally appears to be weak...that described magnetic responses of cryptochrome proteins or organic fluorophores (Karki et al., 2021; Lee... to search for a magnetic response among other proteins that might serve as a starting point for developing...

...fluorescent proteins that are photostable. On top of photobleaching, fluorescent proteins may display ...property of the fluorescent protein. This can differ between fluorescent proteins in the same spectral class...fluorescent proteins? Measuring photostability Since it is clear that a photostable fluorescent protein is a ...performed with soluble fluorescent proteins or localized fusion proteins and do not require dedicated plasmids...fluorescent protein. Photostability measurements can be performed on purified fluorescent proteins. To avoid...fluorescent protein can be identified by a head-to-head comparison of several fluorescent proteins of the ...cyan fluorescent proteins using widefield imaging. Our choice for a photostable protein for live cell imaging...

...the efficiency of BV incorporation by an apoprotein and protein expression level. State-of-the-art FPs ...Shcherbakova et al., 2018). For detection of protein-protein interactions, we recommend using NIR split ...reporter to detect spatiotemporal dynamics of protein-protein interactions. Nat Methods 11, 641-644 (2014...near-infrared BiFC reporter for in vivo imaging of protein-protein interactions. Cell Chem Biol 20, 1078-1086 ...naturally monomeric infrared fluorescent protein for protein labeling in vivo. Nat Methods 12, 763-765...Medicine. Several sets of near-infrared fluorescent proteins (NIR FPs) and biosensors have been created recently...Biliverdin chromophore Near-infrared fluorescent proteins (NIR FPs) were derived from natural photoreceptors...

...materials and information,” we love to share lab protocols that we use as we serve the scientific community...community. Here are a few virus-related protocol videos that may help to save you time and improve your experiment... purification.  Check out the AAV Purification protocol page for more detailed instructions. AAV titration...curve, visit the AAV Titration by qPCR written protocol page. Getting started with tissue culture The...tissue cultures. To keep up on all of the latest protocol videos, lab tips, and more video content from ...video series Resources on Addgene.org Find lab protocols on the Addgene website Watch Addgene's video collection...

...membrane protease VII, which reduces proteolysis of the expressed recombinant proteins. lon protease: Strains...Secreted Proteins: The most common purification protocols are designed for soluble, cystosolic protein products...recombinant protein expression. Many challenges can arise when over-expressing a foreign protein in E. coli...recombinant proteins are typically expressed in E. coli engineered to accomodate high protein loads using...designated lon or Δlon) similary reduce proteolysis of the expressed proteins. hsdSB (rB- mB-): These strains ...trade-off between growth rate and protein quality. For many proteins, a rich media such as TB or 2XYT ...will review the potential pitfalls of recombinant protein expression and some of the most popular commercial...

...PUF proteins! A newly available PUF Assembly Kit makes it possible to devise RNA binding proteins to hit...applying the Golden Gate cloning method to human proteins known as Pumilio/fem-3 mRNA binding factors (PUF...University of Illinois at Urbana-Champaign. The PUF proteins include eight modular repeats, each of which binds...changing amino acids, it’s possible to produce a protein that binds an alternate sequence. By comparison...tagged. “This tool makes it possible to make the protein specific to any endogenous RNA without tagging,...eukaryotic RNA.” Endless possibilities with PUF proteins The tool she and her colleagues led by Zhao described...repressor. In principle, however, designer PUF proteins can be fused to other components to lend them ...

...green fluorescent protein, a red fluorescent protein, a fancy new green fluorescent protein, some affinity...plasmids containing the same protein of interest tagged with different fusion proteins for different experiments...Just express the fusion protein and add a specific ligand! A self-labeling protein gives you the flexibility...tags on different proteins of interest (or one self-labeling tag and a fluorescent protein) enables applications...working with purified proteins in vitro, you can even use the same tag for multiple proteins but label each ...function of the target protein, so it’s always good to check that your fusion protein is functional. In some...& Johnsson, K. (2008). An Engineered Protein Tag for Multiprotein Labeling in Living Cells. Chemistry ...

...the function of a protein or its role in a disease, researchers often isolate proteins of interest and ...and transport of the protein within a living cell. Analyzing differences in protein localization and transport...mechanisms and protein function. Getting ready to determine the subcellular localization of your protein of interest...the many Fluorescent Proteins (FP) available for microscopy, by cloning your protein of interest into a ...expressed as a fluorescent protein fusion, it is possible to track your protein of interest from its production...fusion protein, it's important to validate its function relative to the untagged, wild-type protein. You...wild-type protein visualized with a fluorescently-tagged antibody. By employing “marker proteins” which ...

...selecting a bright fluorescent protein and a photostable fluorescent protein. In the third post of this series..., the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity...applications in which the fluorescent protein is used as an inert protein module. This is explained in more...assayed with purified protein, reflect the likelihood that fluorescent proteins interact or oligomerize...experiments.  Selecting a non-oligomerizing protein A fluorescent protein cannot be assumed to be non-dimerizing...guarantee that using this fluorescent protein to tag your protein of interest will be problem-free. In ...you try a couple of different fluorescent proteins when protein tagging is the goal (Cranfill et al, 2016...

...endogenous protein tagged with a fluorescent protein and repeating this with another fluorescent protein, a ranking... of fluorescent proteins, how to do a head-to-head comparison of fluorescent proteins and how to make ...best fluorescent protein for your application. Selecting the best fluorescent protein for a specific application...task. Many fluorescent proteins are available and the number of fluorescent proteins is steadily increasing... of fluorescent proteins on websites. To make the best choice, the fluorescent protein properties need... fluorescent protein properties and provides a quick way to compare fluorescent proteins, e.g. see Chudakov...reference protein that is translated from the same open-reading frame and separated from the protein of interest...