We narrowed to 695 results for: prot

...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 ...

...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...

... 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...

...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...

...allow one to monitor compartment/protein dynamics or evaluate protein turnover rate. In many cases, however...Fluorescent Protein Plasmids and Resources Relevant Blog Posts Plasmids 101: Green Fluorescent Protein Which...studies or make a reporter construct. Fluorescent proteins (FPs) are also the foundation of multiple important...so thoroughly characterized, it turns out this protein has a few more secrets - during a collaboration...biological research. Photoconvertible fluorescent proteins Newly developed photoconvertible FPs like PsMOrange...repeat the process with these new photoconvertible proteins. With Sattarzadeh et al.’s discovery that GFP ...observed. Sattarzadeh et al. found that only the protonated form of GFP is photoconverted, so this technique...

...molecule, the antibody binds to a protein, and then voila! Wherever your protein of interest is, a signal will...molecules to identify each protein of interest, this gel-based method identifies proteins solely by weight and... isn’t giving you enough signal to detect your protein of interest, it may be time to look at the indirect...antibodies. While a primary antibody binds to the protein of interest, a secondary antibody binds to the ...signaling molecules associated, indirectly, with each protein of interest.    Figure 1: The direct ... molecules are represented in light green; and proteins are represented in orange. Created with BioRender.com...three, secondary antibodies are indicated when the protein of interest is found in low amounts or if the conjugated...

...removes radio-labeled short-lived proteins primarily via proteasome degradation. Autophagy is then induced...supernatant (this is indicative of protein degradation). To control for protein degradation due to other pathways...an active form of the protein. To monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 ... Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors Keima, ...studying autophagy Biochemical assays Long-lived protein degradation: This approach measures the degradation...degradation rate of radio-labeled long-lived proteins as a proxy for autophagy. Cells are cultured with isotope-labeled... to giving more fine-grained detail about what proteins or organelles are being degraded) and it is slow...

... to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one...system.   See our protocol pages for full Lentivirus Production and Transduction Protocols   Once purified...Most of the protocols you will find will recommend using a polycation, like polybrene (protocol) or DEAE ... psi signal, the genes encoding the structural proteins could not be integrated into the new viral particles...plasmid), and 3) the vesicular stomatitis virus G protein (VSV-G) that improves the stability and broadens... one with the exception that the HIV accessory proteins that are not essential for the production of the... to be taken especially if you are working with proto-oncogenes or with human samples that have not been...

...particular the roles of proto-oncogenes, tumor suppressors, and their protein products. Proto-oncogenes consist...refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded by a family of proto-oncogenes. Ras proteins are special...consist of a group of genes/proteins that, when expressed, normally encourage cell survival, growth and proliferation...dividing out of control. One may compare the roles of proto-oncogenes and tumor suppressors to the accelerator...respectively. In the case of a mutation that turns a proto-oncogene permanently “on”, (now an oncogene), this...and in the right places. Checks and balances: protecting against cancerous cells Luckily, there are several...several safety mechanisms in place to protect us from rogue cells when something does go awry. The process...

...labs already familiar with IHC/ICC protocols. A typical ExM protocol (Fig. 1) follows this structure: ...steadily growing in popularity with an abundance of protocols and examples of applications from across disciplines...several times its original size. Depending on the protocol, samples can be expanded to ~4-10x linearly! This... using more accessible imaging equipment, ExM protocols utilize relatively inexpensive reagents and are... Steps 2-4 are not found in standard IHC/ICC protocols and you might expect them to dramatically limit...the types of labels you could use. But many ExM protocols are compatible with conventional antibodies, chemical...chemical dyes, and genetically encoded fluorescent proteins (Gambarotto, et al. 2019; Tillberg, et al. 2016...

...codons that encode a protein may affect the rate of translation and thus protein expression. In fact studies...efficiency and thus protein expression without altering the sequence of the protein. As DNA synthesis has...expressing functional proteins in hosts that do not naturally express that gene. The protein you want to express...detrimental results to the protein. Codon optimization could affect protein conformation, folding and ...strategy for increasing protein production. If you are trying to express a protein in its native host there... Why is codon usage important? Translation and protein synthesis relies on transfer RNAs (tRNAs). tRNAs...cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this study, codon...

...nontarget strand. Protein Cas: Different CRISPR systems involve different proteins, but many of them ...most famous of these proteins is Cas9 nuclease, but there are many other Cas proteins as well, both natural...recognized by the Cas9 protein from the appropriate species. Some Cas proteins, like LbCas12a or AsCas12a...you. We’ll cover the common terminology for the proteins, DNA, and RNA used in CRISPR.   Figure...tracrRNA (in some systems including Cas9), Cas proteins, and a CRISPR array. The CRISPR array is composed...spacer. In bacteria, these crRNAs are bound by Cas proteins (Cas9 shown here). The repeat sequence base pairs...natural and engineered. What these proteins all have in common is the ability to use an RNA guide to target...

...Rep proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While... four Rep proteins share a common helicase and ATPase domains. To function, the Rep proteins form oligomers...viral capsid proteins VP1, VP2, and VP3, which together form the outer capsid shell that protects the viral... start sites generate the three VP proteins. The three VP proteins assemble into capsid at a 1:1:10 ratio...Assembly-Activating Protein (AAP) and the recently discovered Membrane Associated Accessory Protein (MAAP) from...the adenoviral helper plasmid which contains the proteins needed for the virus to replicate,  and the transfer...Replication (rep) gene The rep gene encodes four proteins that are required for viral genome replication...