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We narrowed to 858 results for: HA

Showing: 141 - 160 of 858 results

  1. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ... how a plasmid has been used by multiple labs and see what experimental systems it has been validated ...validated in.  If a plasmid's Addgene ID # has been referenced in other publications, you'll find a link to the... and the PubMed ID # (PMID) for the article that has used this plasmid. Happy researching! If you need...

  2. Hot Plasmids - May 2022

    Type
    Blog Post
    ...   By Ashley Waldron Proximity-dependent labeling has become a powerful approach for proteomic mapping ...our 2019 overview). Until recently, the approach has been hampered by the need to tag each protein of ...M-CREATE) by: Brian O'Neill  The Viviana Gradinaru Lab has previously developed the multiplexed version of the...sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed a method using dCas9-SSAP to knock-in long... methods for large inserts. However, this system has a much lower error rate. For a one kilobase insertion... its functions. Recently, Ravid Straussman’s lab has developed visual barcodes for clonal multiplexing...

  3. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    ...An antibody that has been cited thousands of times may be a safer bet than one that has never been used...work. Whenever possible, choose an antibody that has been validated in your specific application. Consider...cross-reacts with. Ideally you will find an antibody that has been validated in the species your protein of interest.... Talk with your peers. If someone in your field has used the antibody before you will not only have peace...Uhlen, 2016). Be sure that the antibody you choose has been validated using one or more of the proposed ...

  4. Hot Plasmids - February 2022

    Type
    Blog Post
    ...targets by: Mike Lacy Adding to their set of anti-HA frankenbodies (Zhao et al. 2019), Tim Stasevich and...iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag...Adapted from Liu et al. 2021. Anti-FLAG and anti-HA frankenbodies have a variety of applications, including...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs...requires a small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit...larger expression requirements (Figure 1), but Cas12a has generally been underutilized in genetic screens because...

  5. 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...

  6. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...Service since 2009. He has  developed a proprietary, low cost cloning protocol that he has used for cloning ...least 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region... most of the troubleshooting regarding this step has to be strategy specific, there are few general parameters...ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to the strategy adopted...satellite colonies and they form when all the antibiotic has been used. Don’t worry too much, just pick the big...

  7. Addgene is Stopping Use of X (formerly Twitter)

    Type
    Blog Post
    ...communities and welcome suggestions in the comments.  X has moved significantly away from the functionality and...conversation in the scientific community. Addgene has been considering a gradual move off X as a response...conflict with our values. For this reason, our timeline has accelerated. We significantly reduced posting on ...

  8. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while GeoCas9 has maximum activity from 50...the recent discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR...stability in human plasma The last trick GeoCas9 has up its sleeve is its increased longevity in human...stable when incubated in reaction buffer alone, but has limited activity when incubated in diluted human ...

  9. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...You will also have to make sure that your plasmid has been incorporated into the host organism, usually..., c-Flag pcDNA3, comes from the pcDNA series. It has the CMV promoter for mammalian expression, a multiple...mammalian selection cassette (e.g. 1435 pSG5L Flag HA). You can also upgrade your basic mammalian expression...considered to be a more robust expression system and has its advantages; however, remember that you will need...the specific mutations. For these purposes Addgene has a large collection of yeast expression vectors with...

  10. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...until 2005 [1]. Like other transposons, piggyBac™ has two components, a transposon and a transposase. The...offers a large cargo-carrying capacity (over 200 kb has been demonstrated) with no known upper limit [1]....particularly useful when the gene being expressed has threshold effects, such as with signaling molecules...transgenic cell line with the piggyBac™ system is that it has a preference for integration at regions of open chromatin...removal, which alters the endogenous DNA sequence and has been suggested to cause cytotoxicity [5]. Second,...efficiency. While the efficiency of excision with PBx has been shown to be higher than that of the wildtype...Cre recombinase, the PBx is highly specific, and has not been shown to cause genomic abnormalities when...

  11. 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...

  12. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector that can be ...and the subsequent biochemical characterization, has enabled scientists to engineer a wide variety of ...green to red when it is oxidized. The Timer tool has already been used by several groups to monitor protein...imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-... for protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking....optimization through engineering, Ed Boyden’s lab has identified a variety of new channelrhodopsins for...nicknamed Chronos, is blue and green light-excitable and has faster kinetics than those of other channelrhodopsins...

  13. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...between a cell that has had one viral particle integrated into its genome and a cell that has had multiple integration...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected... be significantly higher than the same prep that has been stored in the freezer. If you plan to freeze..., you may want to consider using an aliquot that has been subjected to a freeze/thaw cycle for titering...CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage Tracing Additional Resources...

  14. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...science has become increasingly restricted to academic and industrial labs, but recently there has been ...KittyBiome has been incorporated as a non-profit entity and so far most of their funding has come from ... the predictions about the effects this mutation has on locomotion in our computer model to the effects...Genetics Center (CGC) at the University of Minnesota has declared our makerspace an educational institution...

  15. 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...

  16. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...in living cells. The HA frankenbody is an HA tag detection probe. It contains an HA-binding scFv that’s...often genetically fused to other proteins, which has led to their application for basic and translational...fluorescence intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged...s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe...of cytotoxic granules from the T cell. When a CAR has an scFv that binds cancer-related antigens, binding... Addgene blog: Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more...

  17. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    ...have the biggest impact. We should know — Addgene has built its foundation on this idea! And we believe... cell populations and regions of the brain. This has culminated in a collection of AAV enhancer plasmids... to express exogenous genes — a feature the AIBS has capitalized on. Figure 1: Schematic of transcriptional...Allen Institute’s Genetic Tools Atlas (GTA). The GTA has a wealth of information and includes annotated images...target a very specific cell type. However, Addgene has partnered with the AIBS to showcase the “best-in-...More from the Allen Institute The Allen Institute has many other projects outside of the enhancers and ...research is conceived, developed, funded, and shared has no doubt changed drastically over the past few decades...

  18. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...also the fastest test that has been developed so far.  A similar method has been optimized to detect the...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in...State University. Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting...full-fledged pandemic. Containing the spread of this virus has become an utmost priority for countries around the...the outbreak ultimately flattening the curve, it has become evident that widespread testing is crucial... 83 known COVID-positive samples tested, DETECTR has a positive prediction rate of 95% and a 100% negative...

  19. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ... dark but dissociate under blue light. This tool has now been modified by Torsten Wittmann’s lab at UCSF...genetic or pharmacological manipulations on cells, and has potential use for a wide range of applications in...monomeric, 4-5 fold brighter than mNeptune684, and has a favorable pKa of 5.6. To test out this new FP in...The increasing popularity of single base-editors has resulted in the rapid expansion of the genome editing...expansion of genome editing tools, the Reuben Harris lab has deposited a reporter plasmid that allows for rapid...

  20. CRISPR in the Clinic

    Type
    Blog Post
    ...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...way. Delivery, especially for systemic approaches, has its own unique set of obstacles. Thus far, several...delivery tends to be trafficked to the liver. Thus, it has been used for hATTR therapy, since TTR is primarily...
Showing: 141 - 160 of 858 results