We narrowed to 695 results for: prot

...your protein in the cell or purify it – and you do not have a specific antibody against the protein available... you can always add an epitope tag or a fusion protein. Mammalian vectors One of our most popular mammalian...The addition of an in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo (try pcDNA3...6xHis or GST) to affinity purify the expressed protein with a specific resin (e.g. 6xHis containing pDEST-HisMBP...pDEST-HisMBP). After purification the protein can be released from the resin and used for various binding... collection of vectors carrying several fusion proteins. Pick your promoter, selection cassette and visualization...pattern that allows the function of the encoded protein to be assayed (e.g. pPD95_86 and pPD57_56), or ...

...Viruses are simple: a genome packaged in a protein shell (Taylor, 2014). They’re so simple that we can...own machinery to replicate the viral genome and proteins. This is, for the intrepid researcher looking ...structure While all viruses have a genome inside a protein shell known as a capsid, some also have a lipid...can either follow central dogma (gDNA to RNA to proteins) or require an intermediary DNA replication step...transcribed into mRNA, directly translated into proteins, or, in the case of retroviruses, reverse transcribe...transcribed into mRNA before being translated into proteins. RNA viruses replicate their genomes using a variety...ability. The remaining particle, which retains the protein coat (and envelope), has a “gutted” genome with...

...and using RNA scaffolds to assemble proteins in vivo. Nature protocols. 7(10) 1797-1807 (2012). PubMed. ...CRISPRi) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...into a secondary structure that can bind specific proteins via aptamer domains. Combined with dCas9, this...enzymes for violacein, a distinctive pigmented protein. By altering the transcriptional programs, the ...scRNAs all individually utilize the same dCas9 protein. By using an inducible promoter to modulate dCas9...cell fate decisions. By using an orthogonal dCas9 protein that will still recognize guide RNA but with a ...effectively change metabolic flux. The dCas9 protein and orthogonal variants can be used as regulatory...

...DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol ... plasmid DNA from bacterial cultures is a core protocol in molecular biology. And while kit-free plasmid...quality/low quantity) – Bacterial transformation protocols require such small amounts of DNA (often on the...process and include things like genomic DNA, RNA, proteins, salts, ethanol, or endotoxins. The most common... kits — which to choose? Plasmid purification protocols can be broken into a few broad stages: lyse the...environmental impact. Using non-silica-based purification protocols requires less plastic and avoids the use of more...have suggested modifications to their normal kit protocols.  Yeast plasmids – If you’re extracting plasmids...

...separate enzymes that are proteolytically processed from the larger protein: PR (protease), RT (reverse transcriptase...their life cycle to continue. They rely on a viral protein called reverse transcriptase to convert their RNA...typically encodes the vesicular stomatitis virus G protein (VSV-G), which facilitates viral entry into the... integrated into the host genome via the viral proteins packaged during production (read our blog post...into a large complex of both viral and cellular proteins and trafficked to the nucleus. Since the integrase...transgenes/shRNAs If you want your cells to express a protein or nucleic acid for only a limited time, IDLVs ...Lentiviral Guide and our lentiviral production protocol.  One thing to keep in mind is that titering these...

...fluorescence intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged ... are often the building blocks for engineering proteins. So let’s get acquainted with scFvs. What is an...antibody.  The ability to combine scFvs with other protein domains, much like lego pieces, makes them better...antibodies to be used as building blocks for engineered proteins. This flexibility is largely due to scFvs being...antibodies. scFvs are often genetically fused to other proteins, which has led to their application for basic ... HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just like... Whitlow M (1988) Single-chain antigen-binding proteins. Science 242:423–426 . https://doi.org/10.1126...

... purification protocol involving iodixanol gradients. The current purification protocols for isolating...accumulate inside the producer cells and purification protocols often describe a process that begins with cell... also contain other cellular molecules such as proteins or nucleic acids. This conjunction of vectors ...Since the development of the exo-AAV isolation protocol by Maguire and colleagues, other researchers have...cells also contain cellular macromolecules (e.g. proteins, nucleic acids) which possibly improve transduction...association of AAV particles with membrane vesicles protects them from antibody recognition and inactivation... contexts, such as the delivery of therapeutic proteins to the brain which requires crossing the blood-brain-barrier...

...of a protein, a yes-no plot is the best-case result. For example, you have introduced a GFP protein into...adjusted voltage, cells that did not receive the GFP protein are most likely to be clustered at signals below...not! When you are looking to study the level of protein expression, instead of its presence or absence,...These smears tend to occur when you are looking at proteins that are expressed regularly in cells and might...The orange peis the baseline expression of the protein of interest. Created with biorender.com. The... the orange peak is a baseline expression of a protein while the green (top) has an upregulated expression...Priyamvada Prathima has a research background in protein mutagenesis and cancer immunotherapies. She is ...

... Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated guide RNA...using long ssDNA donors.” Nat Protoc. 13 (2018) (1):195-215. DOI: 10.1038/nprot.2017.153 PubMed PMID: 29266098... example, large insertions, like a fluorescent protein knock-in, suffer from low efficiency when a standard...desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region using primers...the commonly used cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc...You can also use Addgene plasmids to purify Cas9 protein. The table below contains some of our favorites...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...

...editors that can be reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme....Genetically encoded voltage indicators (GEVIs) are proteins that detect and output the voltage changes in ...inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight amino acid...generated knock-out and knock-in mutants, determined protein expression levels from different promoters, and...half with PhyB or PIF6, phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In the presence... light or PCB is no longer present, the fusion proteins separate, inactivating Cre. This is critical for...truncated Phytochrome B (PhyBΔ) and the active protein binding domain of Phytochrome B interacting factor...

...be transformed into Alphaproteobacteira, Betaproteobacteria, and Gammaproteobacteria species isolated from...self-assembling protein (SAP) such as a bacteriophage shell protein or a bacterial microcompartment protein. The ...fluorescent proteins into one particle within the cell, the lab fused the red fluorescent protein (RFP) to... (DCDC), relies on the assembly of fluorescent proteins into one bright particle that segregates into ...

...polymerase (Pol) for replication and Gag structural protein on yet a third plasmid (packaging plasmid). Viral...viruses also have a propensity to integrate into proto-oncogenes, which can lead to cancer. These viruses...viral handling safety   Containment and personal protection are key to lab safety and the same principles...department if they are not currently provided. Eye protection or face shields are required whenever there’s...splash or spray risk in BSL-1. In BSL-2 labs, eye protection is required any time you have to work with BSL... cabinet. Many researchers choose to wear eye protection when they are working in a biosafety cabinet,...organization. It’s always okay to add another level of protection! When centrifuging viruses (a necessary step...

... they bind to a class of Argonaute proteins. The Argonaute proteins remove one strand of the siRNA and...from the Sánchez lab describes the plasmids and protocols required for both WISH and RNAi experiments, as...class” which provides more detailed materials and protocol resources. Find Sánchez lab Plasmids Visualizing...tissue. Once the riboprobes are created, the WISH protocol involves preparation of the tissue sample, several... the translation machinery. In the Sánchez lab protocol, dsRNAs are delivered by feeding them to planarians...which planarian species one is working with, the protocol will slightly differ. For example, Girardia sp...riboprobes and dsRNAs into PR-T4P. The Sánchez lab protocol also provides step-by-step instructions for the...

... this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin dependent manner....a group discussion on the best strategies and protocols for different experiments.  In this blog post...structures, alternative polyadenylation for all protein coding mRNAs, and annotations extracted from other... discovered in plants to conditionally deplete proteins of interest in C. elegans (Zhang et al., 2015)...scientists to both spatially and temporally knock-down proteins of interest.  Plasmids for cGAL and split cGAL...each half to a gp41-1-N-intein. Split inteins are protein domains that naturally “associate, self-excise,...degradation (AID) system enables versatile conditional protein depletion in C. elegans." Development 142.24 (2015...

...the action depends on a surface glycoprotein – the only viral protein present on the virion surface – ...States while caring for a patient, despite wearing protective gear, and now comes news of a second Texas healthcare...does all of its damage with just seven structural proteins. But, as NPR explained, it’s not the virus itself...in PLoS Pathogens shows that the Ebola virus glycoprotein remains essential in allowing those infected...in a novel way, by shielding host cell surface proteins. Researchers at the University of Virginia School...Medicine have also recently described how the glycoprotein transforms itself to escape vesicles within ...reported in the Journal of Virology.  The EBOV glycoprotein’s role in infection also makes it a critical ...

... 2016). The protein capsid encapsulating the genome is majorly composed of three proteins that can be ...of terminal proteins capping the ends of linear double-stranded AdV genomes. Such protein-DNA structures...that AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed...along with a description of their experimental protocol. The three plasmids which have been deposited ...steatohepatitis cancer. In deficiencies of plasma proteins where Adv vector delivery is often used, the CRISPR...producer cells could produce corrective amounts of protein for any such disease. This technology therefore...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK....

...specificity, they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish...applicability of base editing through protein engineering and protein delivery. Nature Communications, 8(...requires three elements. They are, broadly: A Cas protein (catalytically dead or nickase) fused to a deaminase...within the editing window specified by the Cas protein These elements were the starting point towards...efficiency of ~50% and confirmed that the fusion protein retained the PAM properties of the individual Cas9...formation 1.5-2 fold, the team fused bacteriophage protein Gam to the N-terminus of BE4 (Komor et al., 2017..., ABE8s resulted in 1.5-fold more editing at protospacer positions A5-A7 and 3.2-fold more editing at ...

...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...swapping of fluorescent proteins and multiplexing is simple when fluorescent proteins with limited spectral...FRET reporters are comprised of two fluorescent proteins with very specific spectral properties that limit... and the orientation of the donor and acceptor proteins, which can be difficult to predict. Slower Dephosphorylation...single cells using translocation reporters. Nature protocols, 13 1, 155-169. PMID: 29266096. Find the KTR plasmids...biosensors Learn about using phosphoserine to study protein phosphorylation Additional Resources on Addgene.org...

...the methods in this blog post. 8. Chromoproteins: Colorful Proteins For Molecular Biology Experiments ...Experiments by Jennifer Tsang Chromoproteins are a subset of the fluorescent protein family and they generate color...detection methods, synthetic nanobodies, SARS-CoV-2 protein expression, and more. We also highlighted reagent...features 210 plasmids: empty backbones, fluorescent protein tags, SARS-CoV-2 plasmids, etc. It is our hope ...Without A Kit by Leah Schiesow We added to our protocol collection this year and along with it, wrote ...subset of these tools for plasmid mapping, DNA/protein databases, DNA sequence manipulation, and primer...

...Control Protein Activity Learn more about protein tags Read about E. coli strains for protein expression...the bdNEDP1 protease, thus releasing purified nanobodies from the column. bdNEDP1 protease can be generated...expressed in bacteria and extracted with common protein purification methods. Read on to learn more about...be better suited for the job. As single domain proteins, they can be expressed in bacteria, making nanobodies... the bench, this results in a shorter staining protocol because the primary antibody can be “labeled” ...super-resolution imaging, rapid epitope-mapping and native protein complex isolation. eLife. PubMed PMID: 26633879...