We narrowed to 526 results for: HA tag

...clogging during harvesting. The addition of sorbitol has been shown to increase viral titers by up to 1.8-...10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through a 0.22 μm...such as sorbitol and sodium bicarbonate, sorbitol has been found to increase viral titers by up to 1.8-...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Opti-MEM, Thermo Fisher 31985...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...add 55 mL of heat-inactivated FBS and 5 mL of glutaGRO 11 mL of 200 mM L-alanyl-L-glutamine. Store at 4 ....46 M sorbitol, 7.5 mL HI-FBS, and 7.5 mL 100X glutaGRO, 7.5 mL of 7.5% Sodium Bicarbonate, 7.5 mL 1 M...

...final volume of 1 L and recheck pH to ensure that it has not drifted. Filter the solution through a 0.22 µm...incubate for 1 h at room temperature. After the BCD TFX has warmed in the bead bath, transfer the tubes to the...-188 10% Poloxamer 188, Thermo Fisher 24040032 Glutagro, Corning 25-015-CI BalanCD HEK293 Media, Irvine...HEK293 Media 10 mL 10% Poloxamer 188 40 mL 200 mM Glutagro Do not add selective reagents Store at 4 °C until...BCD Feed 500 mL BalanCD HEK293 Feed 20 mL 200 mM Glutagro Store at 4 °C until use. We suggest preparing ...

...polyclonal cell population, meaning that the transgene has integrated in different locations in the various ...variety of dilutions and pick the population that has the most desirable level of expression. Over time...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ... 55 mL of heat-inactivated FBS and 5mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and FBS...

...genome can occur, although at a low frequency which has been estimated to range from 0.01% to 3% depending... well-characterized and extensively studied, and has a broad tissue tropism. Table 1 gives a summary of...across a broad range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue... tropism towards specific cell types of interest has been to create and test capsid libraries with random...systemic capsids. A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...naturally cross the BBB. The Viviana Gradinaru lab has developed a number of systemic rAAV vectors that ...preps . AAV retrograde (AAVrg) Directed evolution has also been employed to engineer an rAAV serotype variant...

...immune response has become a disadvantage for the use of rAdV vectors in gene therapy but has, however, proven...response quickly, which has limited their use in therapeutic applications. However, this has also made them the...the human population has been exposed to at least one human AdV serotype and has developed a serotype-... digest patterns to determine that the transgene has been inserted into the adenoviral backbone plasmid...other immune-modulatory molecules. Another strategy has been to use oncolytic rAdV vectors, which are replication-competent... of spontaneous reversion to RCAs, although this has been reduced by providing the E1 gene in trans within... adenovirus (RCA) Mutated adenoviral vector that has the ability to replicate and reproduce within host...

...of antibody and production strategy has advantages and disadvantages compared to each other. Polyclonal...Experimentally, antibodies conjugated to fluorescent tags or other markers can be injected into an animal,...produced for research purposes, some of which take advantage of natural immune system production and many more...as polyclonal. Polyclonal antibodies have the advantage of more robustly recognizing the target protein...proteins are also plasmid-derived. However, a disadvantage to ScFv is their lower affinity for their target...antigen as canonical antibodies. They also have the advantage of being significantly smaller, meaning they can...proteins you can investigate in each panel. An advantage of flow cytometry is the ability to use multiple...

... it is possible that the difference between them has affected the accuracy of the NanoDrop. It is a good...Background Information During several different stages of molecular cloning, it is important to get a ...

...time it has caught it has usually started to spread (metastasize) and turned into Stage 3 or Stage 4 cancer...and treatments such as these has brought a lot of promise to a field that has historically been full of ...size and may have spread while Stage 4 indicates that the cancer has spread from where it originated to...However, as the U.S. Environmental Protection Agency has noted, there is no safe level of asbestos exposure...abdominal cavity. This targeted chemotherapy approach has been shown to be successful in killing many (if not...patients: early detection and immunotherapy.  It has long been known that one of the best ways to improve...standard treatments and early detection, mesothelioma has proven itself to be susceptible to up and coming ...

...Extend DNA for 1 minute at 72°C: The Taq polymerase has an optimal temperature around 70-75°C so this step...changes during a typical PCR. The DNA polymerase has an optimum temperature around 70°C and is the molecule...cations can also be used for PCR-mediated DNA mutagenesis. A higher cation concentration increases the ...

...the target. CRISPR has revolutionized the generation of knockout cell lines which has made this control...multiple stages of the drug discovery pipeline.   CRISPR and drugs in the future While CRISPR has already...In this blog we will review how CRISPR technology has brought drug and genetic screens together and expedited...candidate genes! Disease model generation CRISPR has made the generation of genome edits accessible, fast... have the precise disease genetics. Since CRISPR has been adapted for use in primary cells, many tissue...drug is equally toxic to both lines, it probably has either high toxicity or high off target effects, ...engineered to have many unique capabilities which has advanced what type of edits can be made in the contexts...

...sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting ...genomic loci is possible using CRISPR, and Addgene has a number of tools for this purpose. Epigenetics began...many TALEN-based technologies, the advent of CRISPR has made targeting much easier! With the help of guide...modification at specified loci. Charles Gersbach’s lab has deposited mammalian expression constructs including...BioRender.com. Tet1 demethylase Ronggui Hu’s lab has created pdCas9-Tet1-CD for targeted cytosine demethylation...Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR...DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion...

..., and Tol2), each of which has several different applications (mutagenesis screens, transgenic animals...excision site after cleavage by the transposase. It has a cargo capacity >100 kB, although integration efficiency...efficiency decreases with cargo size. Sleeping Beauty has a close-to-random integration profile in mammalian...hypersensitivity sites. Like Sleeping Beauty, piggyBac has an integration efficiency in human cells similar ...1996). Unlike Sleeping Beauty and piggyBac, Tol2 has a weak consensus sequence for its preferred target...hypersensitivity sites. Tol2 is only active in vertebrates and has lower efficiency of integration in human cells than... is a truncated version of the original Tol2 and has ~3-fold increase in transposition activity  Tol2...

...enzymes on the block? The Cas family of proteins has many useful genome engineering enzymes, each with...to generate a blunt DNA double strand break. Cas9 has been studied for genome engineering purposes far ...far more than any other Cas family protein and has thus benefitted from countless rounds of optimization...effective at wiping out large regions of DNA and has anti-viral application potential as a result. Fast... Cas enzymes outlined below as well. Cas12a also has significant therapeutic promise due to the fact that...to package for therapeutic delivery by AAV, which has strict capacity limitations. New efforts have recently...for efficient, multiplexable RNA knockdown. Cas13 has also stirred up interest for its potential as a viral...

...Addgenies has helped me get a much better idea of what lies ahead. Thus the “Meet an Addgenie” Instagram interview...Director of Product Management Eric Perkins, who has been in many roles in Addgene since he started, said... set up for failure. Listening to Kenkel’s story has helped me realize that is not true at all. Racheal...medical school was devastating. At the same time, she has an entirely new path to discover and gives her time...finance, and while I've done that, my big takeaway has been the impact I can have on an organization beyond...was one of the biggest takeaways from my co-op. It has been so insightful to learn about different career... a few years ago at a CRISPR meeting.Read his Instagram feature here. You also don’t have to go back...

... expressing IgM. IgM has a pentamer structure, which means each IgM molecule has a binding capacity or...function is unknown. It has a monomer structure with a valency of two and has no subclasses. IgG IgG is... appropriate isotype control. An isotype control has the same constant heavy chain domain as your primary...replaces the hinge region in IgM antibodies. IgM has no subclasses and is found mostly in blood. IgD While...on the Addgene blog Learn about the advantages and disadvantages of using monoclonal antibodies Read ...pathogen. Different isotypes also appear at different stages of an immune response. There are three main pathogen...

...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the world...Publication at Addgene Cas9 and NgAgo head to head NgAgo has a number of unique features compared to Cas9. These...compare CRISPR and NgAgo, but it seems that NgAgo has lower potential for off-target effects. In Gao et...characterized Argonaute from Thermus thermophilus (TtAgo), showing that this protein could cleave DNA in ...Unfortunately, Swarts et al. were unable to show TtAgo-mediated cleavage in mammalian cells, likely due...al. searched for Argonaute orthologs similar to TtAgo and PfAgo, another thermophilic Argonaute. They ...

...Resource Center has various plasmids for overexpression and knockdown. Addgene also has a number of insect... transgene ends up (1): Random insertion has the advantage that you can get varying expression levels... to determine in which chromosome your transgene has integrated. This can be accomplished with a few matings... embryos and pick out flies where your transgene has been incorporated into the germline. Outsourcing ... have to map into which chromosome the transgene has integrated and make genetic crosses with balancer...finishing his immunology PhD at Harvard University. He has had nearly a decade of experience working with Drosophila...derivatives suppress TH17 cell differentiation by antagonizing RORγt activity. Nature. 472, 486–90 (2011). ...

...objective performs essentially the same task but has different magnifying powers. When you look at the...microscope stage. If using a slide, you can secure it into place using the metal clips on the stage. Turn ...Turn on the power source and use the stage arm to move the stage so that the light shines onto your sample...Once your sample is in focus, use the stage arm to move the stage (and ultimately your slide) until your... As the name suggests, light microscopes take advantage of the physical properties of light to detect ....com. Base Light Source Condenser and Diaphragm Stage Objective Lenses Focus Knobs (Fine and Coarse) Nosepiece...Slides (or other sample that can fit on a microscope stage) Reagents None needed Procedure Set up your microscope...

...that for the flow plots discussed in this post, voltage has been adjusted appropriately and compensation ...interpretation. Priyamvada Prathima has a research background in protein mutagenesis and cancer immunotherapies...baseline expression of a protein while the green (top) has an upregulated expression (shift to the right). The...overlaps with the baseline suggesting this sample has a comparable expression to the baseline/negative ...adjusted voltage and compensated your colors. Here are some good practice tips on adjusting voltage: During...changing the voltage for the particular fluorophore prior to recording data. Increasing the voltage will shift...decreasing the voltage will shift it to the left. If your signal is too high, lower the voltage to have your...

...approaches, has its own unique set of obstacles. Thus far, several targeted therapies have taken advantage of ...There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing field ...CRISPR Therapies Cancer and CAR-T therapy The FDA has already approved CAR-T immunotherapy for cancer treatment...CRISPR Therapeutics. Data shared from these trials has been outstanding, with almost all patients experiencing...desired edit, but this particular class of treatments has quite the opposite aim: to destroy. One treatment...delivery tends to be trafficked to the liver. Thus, it has been used for hATTR therapy, since TTR is primarily...targeting the diseased allele specifically. Early-stage trials showed a high success rate in clearing misfolded...