We narrowed to 633 results for: gats

... the same name. The tag has a net negative charge with 4 negatively charged and 1 positively charged amino...hemagglutinin. The tag has two negatively charged amino acids for an overall negative charge. This tag has been...common tags, FLAG is the most charged, with 5 negatively charged and 2 positively charged amino acids....localization and solubility, so be on the lookout for aggregation of your protein of interest!   Ready to start...Arribere, J. A., et al., Translation readthrough mitigation. Nature. (2016) 534, 719-723. DOI: 10.1038/nature18308...

...outside their field, the answers were primarily negative. Stupid, overwhelmed, discouraged, tired…words...from AAV Purification by Iodixanol Gradient Ultracentrifugation First, you’ll want to identify the reader...for AAV Purification by Iodixanol Gradient Ultracentrifugation. The reader may be, for example, a graduate...between a gradient tube before and after ultracentrifugation so that a scientist can extract virus from...interfaces between layers are blurred after centrifugation and the syringe needle is inserted bevel-up...profile photo of an iodixanol gradient after ultracentrifugation. The interfaces between layers are muddled...illustrates a gradient tube before and after ultracentrifugation. The viral supernatant migrates through increasingly...

...detected using an unconjugated primary antibody, then visualized using a fluorophore-conjugated secondary antibody...Antibodies - typically, primary antibodies that are conjugated to a fluorophore, to avoid the use of secondary...the signal comes from the antibody-fluorophore conjugate, and is not expressed by the cell itself, there...lot of flexibility in which proteins you can investigate in each panel. An advantage of flow cytometry.... These could be visualized using fluorophore-conjugated antibodies (i.e., immunofluorescence), or by .... These applications are typically used for investigating signaling pathways or developing therapeutics...therapeutics. Experimentally, antibodies conjugated to fluorescent tags or other markers can be injected into an...

...Method Choose from: Gateway cloning, gene synthesis, Gibson cloning, ligation-independent cloning, restriction...Please indicate which growth strain Addgene should propagate your plasmids in: DH5α, NEB Stable, ccdB Survival...Survival, Pir1, or other. Whenever possible, we propagate plasmids in the standard cloning strain DH5α. For...For plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids...choose the Pir1 strain. If your plasmid cannot be propagated in any of these standard strains, choose other...Distribute pending QC. The name of the Principal Investigator and Organization where your plasmids were first...

... antibody is also conjugated to another molecule, such as biotin. Next, a conjugated substrate is added... biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated to the ...protein of interest is then detected by an enzyme-conjugated antibody. Next, its substrate is added and the...instead of detecting your protein using an enzyme-conjugated antibody, a primary-secondary pair is used. This...single primary antibody. The secondary antibody is conjugated to an enzyme for visualization. The indirect ...

... by ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus...vectors) you can concentrate your virus by ultracentrifugation of the viral stock, followed by resuspension...packaging cell debris before proceeding to ultracentrifugation to avoid contaminating your transduction ...through a 0.45 µm or 0.22 µm filter) or by a centrifugation step (5 minutes at 300-500 g). After removing... to concentrate a virus solution without ultracentrifugation, you can reduce the volume of cell culture...repulsive electrostatic forces between these two negatively-charged membranes (Davis et al., 2002). Polybrene...

... a specific conjugate, and changing your reporter molecule requires either re-conjugating or re-purchasing... while providing flexibility in assay design. Conjugated secondary antibodies are relatively cheap and...westerns typically rely on primary antibodies conjugated to fluorescent proteins, instead of the more ...the enzymes used for chemiluminescence can be conjugated to primary antibodies). Direct westerns are simpler...or loading sample, which may not be practical. Conjugated primary antibodies can be more difficult to source...antibody with the new reporter. Finally, each new conjugate will have to be tested to ensure it doesn’t impact...if you prefer the DIY method. As some proteins aggregate at 95 °C, you may want to check for any available...

...all the latest hot plasmid updates!     Tol2 Gateway-compatiable toolbox for studying the nervous system...system The Cole lab recently deposited the Tol2 Gateway-Compatible Toolobox, which will allow scientists...neurobiology and neuorodgenerative diseases. The Tol2 Gateway-Compatible Toolbox is based on the original Tol2kit...empty vectors that have standard cloning sites or gateway compatible cloning sites for easy cloning of your...toolbox adds new neuronal tools to the expanding gateway compatible vectors currently shared amongst the...2007. PubMed PMID: 17937395 Listen to Our Tol2 Gateway-Compatible Toolbox Podcast Segment The impact of...to premature aging and cancer. The Denchi lab investigated an aspect of telomere length regulation called...

...constructs with Golden Gate MoClo. The barrier for those switching to Golden Gate is a conceptual barrier...synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning (MoClo) Kit. What does it include...people already use. It makes the switch to Golden Gate Cloning easier because individual labs will not ...soon to make it easier for people to use the Golden Gate cloning system especially with plants in mind. When... work in cancer biology.  References: A Golden Gate Modular Cloning Toolbox for Plants. Carola Engler...Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...

... fluorophore conjugates are used, but these are usually more expensive than unconjugated primary antibodies...cysteine. Maleimide-conjugated HRP can be found here and several maleimide-conjugated dyes are also commercially... primary antibodies with secondary nanobodies conjugated with the desired fluorophores. In figure 4B below...individually incubated with a secondary nanobody conjugated with Alexa Fluor 488, Alexa Fluor 568, or Alexa...cysteine sites the nanobody has available for conjugation to a specific label (see step 4 for more information...

...restriction enzyme digestion followed by ligation. Upon ligation, the original pattern of the MCS is regenerated...of the ligation product, and the restriction site between the modules is eliminated upon ligation. As a...regenerated at the flanks of each ligation product, which is the basis for the repeatability of the method...added to the 5’ terminus or the 3’ terminus of the ligation product. Comparison to other high throughput cloning... Method Technology Pros Cons References Golden Gate Type II restriction enzymes Can assemble up to 10...

...microscopy after negative staining. Empty vector particles can be identified after negative staining and ...preparations are purified by iodixanol gradient ultracentrifugation and concentrated to purity and titers adequate...final vector preparation, the quantity of gram-negative bacterial endotoxin is ensured to be less than...containing lentivirus is first cleared by low-speed centrifugation and then by filtration through a 0.45 µm polyethersulfone...in 10% polyethylene glycol (PEG) followed by centrifugation. Precipitated pellets containing viral particles...possible, all plasmids used for viral production are propagated in the endA-mutated NEB Stable strain of E. coli...

...alternatives to the standard restriction/ligation, Gateway, or Golden Gate cloning methods. Whether it is for...based technologies We've described Sequence and Ligation Independent Cloning (SLIC) in a previous Plasmids...polymerase extension cloning) and SLiCE (Seamless Ligation Cloning Extract) to name two more. Likewise, there... work by significantly different methods. Golden Gate cloning does result in the seamless joining of fragments...location and NOT in the middle of your insert. Gateway cloning utilizes λ integrase to catalyze directional...

...Positive and negative controls Most biological experiments require positive and negative controls to ensure...experiments will benefit from positive (+) and negative (-) controls in addition to experimental samples...In this case, mouse samples from positive (+), negative (-) and experimental (?) groups were run on a ... check for tau molecular weight shifts. A negative control is a sample or group that is not subjected...experimental condition. In the example above, a negative control could be a third group of mice that is...receive either protein kinase inhibitor. Since the negative control group does not receive the drug, any response...

...fluorescently-conjugated streptavidin to get more fluorophores per antibody, or conjugate with an enzymatic...primary antibody to bind the target, wash, add dye-conjugated secondary antibody that recognizes the primary...do simultaneous or sequential labeling is what conjugated secondary antibodies you have available. If the...only secondary antibodies you have on hand are conjugated with the same dye (say, you have Goat Anti-Rabbit... direct IF, using primary antibodies that are conjugated to fluorophores. However, the primary-secondary...or scFvs. Many, many options for fluorescently conjugated secondary antibodies are commercially available...

...process consisting of digestion, purification, ligation, and transformation. While restriction cloning... developed. Some of these include, TA cloning, ligation independent cloning, TOPO cloning, one step cloning...developed by Li, et al., in 2011. This method is ligation independent, it does not require purification ...Amplification + + Digestion + + Gel Purification + - Ligation + - Transformation + + The FastCloning process... reactions. These sticky ends are annealed and ligated in vivo. Detailed FastCloning procedure To start...highly simplified, purification-free, sequence-and ligation-independent PCR cloning method." BMC biotechnology...

...innovative tools enabling large-scale real-time interrogation and monitoring of brain cells and circuits during...

...components: It's important to assess how dye-conjugation affects protein function and its propensity to...process: The process of dye labeling, such as conjugation with proteins, should be carefully managed. Proper...degree of labeling (DOL), and thorough removal of unconjugated dye molecules are critical for accurate and ...assay results (Hayashi-Takanaka et al., 2014)​​. Conjugates with lower DOL tend to have weaker fluorescence...fluorescent dye. For instance, the cleavage of FITC-conjugated protein ligands is unaffected by additional protein...non-specific binding, and quenching, which can be mitigated through careful selection and optimization. Fluorescent...Evaluation of chemical fluorescent dyes as a protein conjugation partner for live cell imaging. PLoS ONE, 9(9)...

...dye-nitrilotriacetic acid (NTA) conjugates (Kapanidis, Ebright, & Ebright, 2001). These Cy/NTA conjugates work the same ...tag from 6 to 10 residues. While these Cy/NTA conjugates are not commercially available, they are easily...in intact cells. The Gradia Lab has deposited a gateway vector with Addgene, where you can insert your ...adjacent to each other on the same protein and a negative control experiment conducted on donor-only labeled... (Ni2+: nitrilotriacetic acid) n-fluorochrome conjugates." Journal of the American Chemical Society 123.48...Your Experiments Beware of Fluorescent Protein Aggregation Brush up on Your Fluorescent Protein Basics ...

...gRNAs. As each ligation reaction uses a pool of gRNAs as starting material, the ligation products are a... et al., 2014) individual genes for studies interrogating their functions. The continual advancements ...with straightforward restriction digestion and ligation steps to build barcoded lentiviral plasmids containing...lentiviral plasmids. Multiple rounds of digestion and ligation result in plasmid libraries containing combinations... developing and applying new technologies to interrogate and understand complex biological systems. He...