We narrowed to 695 results for: prot

...existed! Addgene recently started automating a few protocols to make standard lab processes easier and more...own lab. Automating bacterial transformation One protocol that has been automated is high throughput transformation... machine that would improve our transformation protocol and, after looking at a few options, we decided... to use the FXP to automate and optimize more protocols in the future - something that will be key for...same amount of time by hand! Setting up a basic protocol on the Biomek FxP is pretty simple. By clicking...perfectly. Working with Qiagen, we developed a protocol to run up to 192 samples at a time. It follows...contamination between samples. With this automated protocol, we can prep two 96 well plates in just under ...

... of neural circuits. Nature protocols. 2012;7(1):12-23. doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972..., most of which is relatively cheap). Several protocols have been written outlining the different steps...steps and materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various...ferrule implant must be maintained during the entire protocol, otherwise optical stimulation will be erratic...generation (TTL pulse) is a standard electronics protocol not specific to lasers or optogenetics so cheaper...Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje...channelrhodopsin for activation of CNS neurons. Current protocols in neuroscience / editorial board, Jacqueline ...

...-F, but instead employ a Cas12k protein instead of three Cas proteins (Fig. 1A, bottom), and require only...spacer sequence specifying the target site. Four proteins (TniQ, Cas8, Cas7, Cas6) that forms the QCascade...module together with the crRNA.  Three transposase proteins (TnsA, TnsB, TnsC). The donor DNA (aka mini-transposon...INTEGRATE. The requirement for multiple Cas proteins to form the Cascade targeting complex is a hallmark...Type I CRISPR-Cas systems, in contrast to single-protein effectors employed by Type II systems (such as ...forms a novel co-complex with the transposition protein TniQ for targeting (Halpin-Healy et al., 2020).... only two transposase proteins (TnsB and C). While requiring fewer components for RNA-guided DNA integration...

...well-established protocols for their production and purification that yield high titers (see also AAV protocols for...Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal tracing compounds... For predominantly retrograde tracing, albumin protein labelled with horseradish peroxidase (HRP; Kristensson...contained by a capsid which is comprised of the viral proteins VP1, VP2, and VP3. To date several hundred variants...engineered to have differences in their capsid proteins. For a brief overview see Addgene’s AAV guide....neuron-specific transgene expression. Fluorescent proteins are widely used for tracing and their expression...that express one of a few different fluorescent proteins (red, green, blue). If a researcher injects a ...

...tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine deaminase for RNA editing... Burkholderia, they identified 67 genera of Proteobacteria that this toolkit might be used in. Read the...engineered SECURE adenine base editor variants using a protein-truncation strategy with reduced RNA-recognition...abilities of C-to-T and A-to-G base editors into one protein. They fused an activation-induced cytidine deaminase...bioRxiv Find the ORANGE toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle...bionanoparticle system for packaging Cas9/gRNA ribonucleoproteins for transient expression. They replaced the...with a com aptamer which binds an aptamer binding protein for packaging into a lentivirus-like particle. ...

...permuted green fluorescent protein (GFP), and nLightR uses a red fluorescent protein, mApple. Kagiampaki et...that red fluorescent proteins tend to be less sensitive than green fluorescent proteins. nLightR did perform...S., & Steinegger, M. (2022). ColabFold: Making protein folding accessible to all. Nature Methods, 19(6...More resources on the Addgene blog Fluorescent proteins 101: Fluorescent Biosensors Which Fluorescent ...Find Optogenetics Plasmids Visit the Fluorescent Protein Guide Pages ...

...setting. Read more: Plasmids 101: Protein Tags Produce proteins that glow so that scientists can track... a plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection...gene is expressed in and amount of recombinant protein obtained. Selectable Marker The antibiotic resistance...used to do include: Produce large amounts of a protein so that scientists can purify and study it in a...inside a cell Plasmids 101: Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the ...plasmid stocks. The guide also contains multiple protocols and troubleshooting tips to make plasmid usage...

...fluorescent protein followed by a stop codon and the coding sequence for a green fluorescent protein (GFP)....biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The reaction...-free system, capable of producing fluorescent proteins and enzymes. Image from Moore et al., 2021. ...through monitoring the expression of fluorescent proteins and oxytetracycline enzymes. Next, this system...libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline...messenger-RBP sgRNA Library targets human mRNA binding proteins with 10 guide RNAs per gene. The lab modified ...

...recombinant proteins at high levels. In a case where clinical applications of recombinant proteins are necessary...to the production of clinical viral therapies. Protein production  HEKs are extremely easy to transfect...eliminate non-human PTMs which is a concern with other protein production systems, such as yeast. HEKs can also...cell line’s ability to faithfully produce these proteins and its relatively low expression of native channels...receptor biology, ion channels, or involve human protein preparations with PTMs. Why the HEK not? References...Cell Line as a Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021...

...structural component of atherogenic lipoproteins, shuttles lipoproteins through the circulatory systems, ...LipoGlo: A reporter for measuring ApoB bound lipoproteins Article contributed by Leah Schwiesow A leading...cardiovascular disease is driven by atherogenic lipoproteins that accumulate within the vascular wall. ApoB...formation of atherosclerotic plaques. ApoB bound lipoproteins (ApoB-LPs) are indirectly measured through their...requires binding of bacterial enhancer binding proteins (bEBPs) at a sequence upstream of the promoter...fused to an RNA binding motif, and a RNA-binding protein-bEFP fusion. Thus, the bEFPs bind the UAS via the...

...concentrating your virus. See our Concentrating AAV Protocol (Step 10) for instructions on how to do this. ...necessary to visualize expression of fluorescent proteins, for example, as many viral particles must infect...infect a cell to achieve visible expression of the protein. However, expression of receptors may not need ...In the case of channelrhodopsins, fewer channel proteins are required for the transduced cell to become...become light responsive than fluorescent proteins required to get a good fluorescent signal. It may not be...take below but you should read our AAV Production Protocol for additional best practices. Keep your cells...Addgene.org Find Ready-to-Use AAV Preps Find Lab Protocols for Work with Virus Watch Our AAV Purification...

... 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...

... 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...

...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, ...

... 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...