Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 520 results for: HA tag

Showing: 241 - 260 of 520 results

  1. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    ...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...

  2. DNA Quantification

    Type
    Protocol
    ... 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 ...

  3. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...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 ...

  4. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...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 ...

  5. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...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...

  6. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...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...

  7. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ..., 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...

  8. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...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...

  9. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...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...

  10. Antibodies 101: Isotypes

    Type
    Blog Post
    ... 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...

  11. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...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 ...

  12. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...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). ...

  13. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...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...

  14. CRISPR in the Clinic

    Type
    Blog Post
    ...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...

  15. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...diverse bacteria from various environments. It has the advantage of stable genomic integration and is easily...Flp recombinases used to excise markers. It has the advantage that CRISPR efficiency can be controlled by...Cas9-adenosine deaminase fusion Caixia Gao’s lab has developed a new plant base editor from an evolved...String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR), developed by Stefan Stricker...high numbers of gRNAs in one overnight reaction. STAgR is highly customizable as vector backbones, and ...chosen. Read the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi...

  16. Using a Light Microscope Protocol

    Type
    Protocol
    ...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...

  17. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...to define how you will know that the relationship has been successful – what are the desired outcomes? ... Dora Farkas, founder of Finish Your Thesis (who has also blogged for us) offers a free report on her ...called “Secrets to Success in Graduate School”. She has also launched an online career coaching program that...in Madison, Wisconsin she also works remotely and has experience as a scientific recruiter. Chris Humphrey...of From PhD to Life. She runs great tweet chats, has an outstanding blog and, while her PhD is in History...remotely, will travel for speaking engagements and has many areas of specialty from leadership development... consultation meeting or call for free.  Take advantage of this to get the information you need to choose...

  18. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ... The key advantage of using RNA to reprogram cells is that it’s simple and efficient. It has a high efficiency... field of induced pluripotent stem cells (iPSCs) has been around for 10 years. In that time, scientists... of Myc or other oncogenes after differentiation has been linked to tumor formation in iPSC-derived and...iPSCs compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared to MMLV-derived...mechanism between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...cell division. A drawback to this approach is it has lower levels of efficiency at generating iPSCs, usually...

  19. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...methods. Each method has its own pluses and minuses, but Golden Gate cloning has been especially useful...of potential clones. Golden Gate assembly has a few advantages over other cloning methods. Exonuclease-...just a few steps. More recently, CRISPR technology has adapted Golden Gate cloning for inserting the appropriate...also be adapted to express multiple gRNAs. Addgene has two Golden Gate-based gRNA assembly methods available...added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning Golden Gate cloning is...destination vector, making multiplexing easy.  Disadvantages of Golden Gate cloning Golden Gate cloning is...

  20. Technical Design of a Western Blot

    Type
    Blog Post
    ...agent than BME and has a less… distinctive smell. BME smells like rotting eggs but has a longer shelf life... It doesn’t degrade during long-term storage and has both high binding capacity and high protein retention... specialty low-background PVDF membrane, it also has higher background noise than nitrocellulose membranes... other hand, doesn’t require any pre-wetting and has lower background than PVDF. It is a little quicker...quicker to use, with high binding capacity, though it has lower retention after binding and washes. It’s less...and need to be read fairly soon after the reaction has started. Types of chemiluminescence enzymes The two...darkroom and a developer. Film is highly sensitive but has a limited dynamic range, which means it may take ...
Showing: 241 - 260 of 520 results