We narrowed to 883 results for: prot

... efficiency >70%. CRMAGE is an incredibly fast protocol, with only 5 hours incubation time needed for ...single round of editing, and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli...components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting module with...with the crRNA, (3) three transposase proteins, and (4) the donor DNA. By using a multi-spacer CRISPR array...system has generally low efficiencies or can be proteotoxic when used in Mycobacterial species, Sarah Fortune...Zalatan’s lab used an approach to identify known proteins involved in the activation of transcription including...including transcription factors, phage proteins, and RNAP components (Dong et al., 2018). They found a...

...-CoV-2 protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins at the...2 N protein  pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 M protein pLVX-EF1alpha-SARS-CoV...months: Lentiviral preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent...2 ORF3a Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) ...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...

...macromolecules used in scientific research such as DNA or protein. It comes as no surprise then that stability of...crRNA (tracrRNA) which interacts with the Cas9 protein. These RNA molecules are vulnerable to attack by... as they will be recognized as foreign RNA. To protect the RNA from degradation, several simple methods...added to the 2’ hydroxyl group of the ribose to protect it from nucleolytic attack. Another stabilizing...gRNA is surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same...enzyme to pair with your guide and our available protocols to get your CRISPR experiment moving. Happy editing...How to design your CRISPR gRNA CRISPR 101: Ribonucleoprotein (RNP) Delivery Components of CRISPR/Cas9 ...

...rest of the protein in place including FokI endonuclease domain as well as fluorescent proteins used to sort...mammalian or other cells. TALENs are engineered proteins that comprise a DNA binding domain and an endonuclease...technology allows you to create a pair of designer proteins that will introduce a double-strand break in the...transfection or electroporation. All you do to create a protein that binds those 15 base pairs is take four plasmids...well-suited for use in stem cells. (See the complete protocol at StemBook). Addgene: How are you now using this...TALEN on paper to a finished plasmid. With our protocol, it is feasible to produce genetically modified...

...engineer the most miniscule parts of life, like proteins, to study or alter their function for a variety...evolution is a method employed to rapidly change a protein towards a specific goal, like improved specificity...antibody engineering technologies, the Institute of Protein Innovation (IPI) provides information on their ...that target families of cell surface and secreted proteins (glypicans, integrins, and more coming soon) as...on using yeast display, structural biology, and protein engineering to develop antibody-based cancer therapeutics...Wittrup, K. D. (2022). Yeast Surface Display for Protein Engineering: library generation, screening, and... (2015). Methods for the directed evolution of proteins. Nature Reviews Genetics, 16(7), 379–394. https...

...antibody. Do you immediately visualize a Y-shaped protein reminiscent of the Addgene mascot Abi? If so, you...When assembled, the four chains make a Y-shaped protein that can be divided into the Fragment Crystallizable... different regions, both through genetic and proteolytic approaches. This comes in handy when, for example...  F(ab')2 - These fragments are produced by proteolytic digestion that retains an antibody’s disulfide... kDa.  Fab - These fragments are produced by proteolytic digestion above the hinge-region, which results...the fragment could bind not only the endogenous proteins you are trying to block but your primary antibody...the antibody you do and don’t need, and how the protein will interact with other components of your experiment...

...antigens: GFP, RFP,  the histone protein H2A2B, and the actin-binding protein gelsolin, but any nanobody sequence...Antibodies are a go-to tool for detecting a protein of interest in cells and tissues. Although antibody... with most multi-labeling immunohistochemical protocols where either a directly labeled primary or a primary...sections Amplifying weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution across...secondary antibody. Compatible with multi-labeling protocols when other antigens are detected with either a...here. Can’t find a RANbody against your favorite protein? No worries! As more nanobodies are developed, ...

... RepA protein under the control of IPTG. Because RepA is a plasmid replication initiator protein, they...using a promoter that is responsive to a repressor protein, the increase in expression due to copy number ...copy number increases, the amount of repressor protein also increases thereby intensifying repression ... from Segall-Shapiro et al., they chose a TALE protein as the repressor because TALEs can be programmed...deoxychromoviridans requires three genes whose proteins are needed sequentially. When Segall-Shapiro et... of products is needed, such as within a multi-protein complex, stabilized promoters can eliminate variability...

... benefit of working with surface proteins is also that such proteins are easily studied in living cells...cause the up-regulation of an important surface protein (e.g. PD-L1 on a cancer cell). In this screen, ...could be targeted. Cells failing to upregulate the protein following stimulation would then be separated by...molecules were involved in the phenotype. Surface proteins  My group works on immune cells in different contexts.... Thus, a screen can be used to target surface proteins to determine which contact points are involved... used to interfere with the identified surface proteins, and such antibodies could thus be interesting...differentially expressed genes that code for surface proteins. One of the first challenges with a custom screen...

...probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering what the...RNAs (ncRNAs) are transcribed but do not encode proteins. They play important roles in many cellular and...functionally manipulate and study in comparison to protein-coding genes (Rinn and Chang 2012). Scientists...Scientists have determined the function of many protein-coding genes thanks to systems like CRISPR/Cas9 that allow...often used to knock-out, or permanently disable, protein-coding genes by generating insertions or deletions...powerful tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription...lncRNAs and has fewer off-target effects. Newer Cas protein variants may also be useful for targeting ncRNAs...

...indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (e.g., calmodulin or troponin...Many calcium reporters use a single fluorescent protein (FP), requiring only a simple measurement. However... mechanism-guided design of single fluorescent protein-based biosensors. Nature chemical biology. 2021...encoded calcium indicators. Cold Spring Harbor Protocols. 2012 Jun 1;2012(6):pdb-top069609. Palmer AE, ...genetically targetable fluorescent indicators. Nature protocols. 2006 Aug;1(3):1057-65. Resources on Addgene.org....org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’s Viral Services  Resources on ...

...tags to the C-termini of Met proteins, which can be used to confirm protein expression by western blotting...function wide open. What if two seemingly similar proteins have acquired divergent functions due to mutations...their experiments, students test whether or not proteins from the fission yeast, Schizosaccharomyces pombe...deposited with Addgene, to successfully analyze Met protein complementation. In the procedure diagrammed in...manuscript in preparation), indicating that Met protein functions have been largely conserved since the...Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...TadA7.10 in ABE7.10)  a catalytically impaired Cas protein (dCas or Cas nickase)  a guide RNA that targets...to ABE7.10. Depending on the position in the protospacer, the editing capacity of the ABE8s ranged from...more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase the targeting scope of adenine base...site in the genomic DNA to contain a specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N... Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG) to create NG-ABE8... genomic editing by measuring the reduction in protein expression of these six genes. All ABE8s edited...efficiently than ABE7.10 with ABE8.20-m reducing protein expression the most in primary human T cells. While...

...figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning method procedure...gene of interest (or, if your protein is fused with a fluorescent protein), fluorescence microscopy can...on Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression... place to screen your clones. Depending on the protein, we have used Western blots, dot blots, ELISAs,...

...fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent protein tools. Future...to creating and sharing many great fluorescent protein tools, Professor Goedhart is very active on Twitter...Resources from the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's Series on Choosing... Choosing the B(right)est Fluorescent Protein Plasmids 101: Visualizing Subcellular Structures and Organelles...Organelles Resources at Addgene.org Find Fluorescent Protein Plasmids Find Plasmids for FRET Find Plasmids for...

...sections: Protocols To complement our written protocol materials, we have also created video protocols.  These...navigate various lab processes.  Our most recent protocol video detailing Streaking Plates for Single Bacterial...Check Our Our Protocol Pages Browse our Plasmid Reference Pages ... protocol video...

...Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm...below affect continuous cellular processes (e.g., protein synthesis), while amp only affects the cell wall...Kanamycin, or kan, is an aminoglycoside which inhibits protein synthesis. The kan resistance gene, NPTII, facilitates...streptomycin Spectinomycin and streptomycin both inhibit protein synthesis as their mode of action and have a very...gene provides zeocin resistance by expressing a protein that directly binds to zeocin, preventing it from...Addgene.org Addgene’s Basic Molecular Biology Protocols Over-Agar Antibiotic Plating Plasmids 101 eBook...

...adenovirus type 5 (hAdV-C5) can be considered the prototype, although many viruses infecting humans (from ... the nuclear compartment, gene expression, and protein translation. Its genome consists of “early” (E)...Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole process – such...absent, and the late genes (and their structural proteins, which would encapsidate new virions) are not ... is transient due to the fact that AdV surface proteins (especially fiber’s knob domain, and hexon), as... as well as other intracellularly-exposed proteins and downstream cellular events, all recruit strong ...J., & Wold, W. S. (1996). The adenovirus death protein (E3-11.6K) is required at very late stages of infection...

...optogenetics — and shared the protocols we use in our own labs. By 2023, we had 47 protocols and 12 guides available...experiment.     Table 1: Most popular guides, protocols, and collections of 2023. Educational Resource...Guide Optogenetics Guide γ-Retrovirus Guide Protocols Agarose Gel Electrophoresis Bacterial Transformation...around the world. Collectively, our 12 guides, 47 protocols, and 186 collection pages received over two million...a thousand words, especially when it comes to protocols. In 2013, we launched the Addgene YouTube channel...Figure 4). Four of the five most popular videos are protocol videos, where showing exactly how to execute a...

... of proteins was mutated, as reported in various studies. We made a table that listed each protein in ...then for each protein we listed all the studies that reported mutations in that protein (including how...because you could easily see how frequently each protein was mutated, you could see how big the studies ...Another example: we made a schematic of all the proteins in the class that showed the relative sizes and...sense of the similarities and differences among proteins in the class. Offer your perspective It doesn...